A peptide used for immunotherapeutics

ABSTRACT

Provided is a peptide provided herein includes at least one peptide unit, and the peptide unit may include at least one B-cell epitope, at least one Th epitope, and an appropriate number of auxiliary parts. The peptide unit is a portion designed to uniformly induce only the intended antibody while exhibiting a certain level of immunogenicity in the body of a subject. In addition, the peptide unit is designed with a relatively short length, and thus has the characteristics of easy synthesis and a low production cost. The peptide has properties suitable for use as an immunotherapeutic due to the characteristics of the peptide unit described above. In the present specification, the design principles of the peptide and the peptide unit are disclosed in detail.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a § 371 national-stage application based onPCT/KR2021/009453, filed on Jul. 21, 2021, which claims priority fromKorean Application 10-2020-0091031, filed on Jul. 22, 2020; KoreanApplication 10-2020-0091032, filed on Jul. 22, 2020; and KoreanApplication 10-2020-0091033, filed on Jul. 22, 2020. The entire contentsof PCT/KR2021/009453 are incorporated herein by reference in theirentirety.

INCORPORATION BY REFERENCE OF SEQUENCE LISTING

The instant application contains a Sequence Listing which has beensubmitted electronically in ASCII format and is hereby incorporated byreference in its entirety. Said ASCII copy, created Oct. 7, 2022, isnamed “PYH-00901_Sequence Listing” and is 156,655 bytes in size.

TECHNICAL FIELD

The present disclosure relates to a peptide that is injected into thebody of a subject to generate humoral immunity, which is a technology inthe field of immunotherapeutics.

BACKGROUND ART

The purpose of an immunotherapeutic is to be introduced into the body ofa subject to induce humoral immunity against the immunotherapeuticitself, and to thereby treat a specific illness or disease with theantibody produced as a result. In particular, the “treatment” alsoincludes prevention of a specific illness or disease. Immunotherapeuticsare similar to vaccines in that they induce antibody production throughan antigen-antibody reaction in the body of a subject; however, theydiffer from vaccines in that the antibody has not only a binding abilityto the immunotherapeutic itself, but also a binding ability to aspecific target in the body (e.g., specific tissues and cells in thebody, or substances generated in metabolic processes, etc.) therebymaking it possible to treat a specific illness or disease, andadminister repeatedly.

DISCLOSURE Technical Problem

The present disclosure provides a peptide which has a function ofinducing a pre-designed antibody in the body of a subject.

The present disclosure provides a composition for an immunotherapeuticincluding the peptide.

The present disclosure provides a nucleic acid sequence encoding thepeptide.

The present disclosure provides uses of the peptide and a compositionfor an immunotherapeutic including the peptide.

Technical Solution

According to an aspect of the present disclosure, there is provided apeptide unit (a block of peptide) which is 23mer to 71mer in length, isrecognized by CD4+ T-cells to induce a humoral immunity, and includesthe following:

at least one Th epitope, wherein the length of the Th epitope is 8mer to32mer; and

at least one B-cell epitope,

wherein the B-cell epitope is a fragment or a mimotope of apolipoproteinB-100, which can induce an antibody targets apolipoprotein B-100.

In an embodiment, the peptide unit has a length in the range of 26mer to50mer, and the Th epitope has a length in the range of 11mer to 13mer.

In an embodiment, the peptide unit includes one B-cell epitope and oneTh epitope, and the peptide unit has a length in the range of 26mer to45mer.

In an embodiment, the peptide unit includes one B-cell epitope and twoTh epitopes (which are each referred to as a first Th epitope and asecond Th epitope); the peptide unit has a length in the range of 37merto 50mer; and the first Th epitope is linked between the B-cell epitopeand the second Th epitope.

In an embodiment, the peptide unit includes two B-cell epitopes (whichare each referred to as a first B-cell epitope and a second B-cellepitope) and one Th epitope; the peptide unit has a length in the rangeof 45mer to 50mer; and the second B-cell epitope is linked between thefirst B-cell epitope and the Th epitope.

In an embodiment, the peptide unit includes two B-cell epitopes (whichare each referred to as a first B-cell epitope and a second B-cellepitope) and one Th epitope; the peptide unit has a length in the rangeof 45mer to 50mer; and the Th epitope is linked between the first B-cellepitope and the second B-cell epitope.

The present disclosure provides a nucleic acid encoding the peptideunit, or a peptide that does not include a nonstandard amino acid amongthe peptides.

The present disclosure provides a peptide in which 2 or more and 5 orless peptide units are linked.

The present disclosure provides a pharmaceutical composition fortreating obesity including the following: the peptide unit or thepeptide; and adjuvants.

The present disclosure provides a method for treating obesity includingthe following: administering the pharmaceutical composition into thebody of a subject.

The present disclosure provides a use of the peptide unit or the peptidefor treating obesity.

The present disclosure provides a use of the peptide unit or the peptidefor preparing a therapeutic for obesity.

Advantageous Effects of Invention

When the peptide provided in the present disclosure is injected into thebody of a subject, the peptide has the effect of inducing the productionof an antibody with a specific physiological function, whichspecifically binds to a previously designed antigenic site.

BRIEF DESCRIPTION OF DRAWINGS

FIGS. 1 to 3 , which show the results of the peptide effect confirmationexperiment according to Experimental Example 2, are graphs showing themeasured changes in body weight per week according to each experimentalgroup.

FIG. 4 , which shows the results of the peptide effect confirmationexperiment according to Experimental Example 3, is a graph showing themeasured weight per week of age of a test subject for each experimentalgroup, in which Lean denotes a control group with normal weight, Obesitydenotes an obesity group induced by high-fat diet, Mock denotes a groupadministered with placebo, and 3H-OTP denotes Group 2-1.

FIG. 5 , which shows the results of the peptide effect confirmationexperiment according to Experimental Example 3, is a graph showing themeasured antibody titer observed in a test subject for each experimentalgroup, in which Obesity denotes an obesity group induced by high-fatdiet, Mock denotes a group administered with placebo, and 3H-OTP denotesGroup 2-1.

FIG. 6 , which shows the results of the peptide effect confirmationexperiment according to Experimental Example 3, is a graph showing themeasured lipolysis ability of hormone sensitive lipase in adipocytes ofa test subject for each experimental group, in which Basal denotes acase where norepinephrine was not treated and Hormone denotes a casewhere norepinephrine was treated, and Lean denotes a control group withnormal weight, Obesity denotes an obesity group induced by high-fatdiet, Mock denotes a group administered with placebo, and 3H-OTP denotesGroup 2-1 ((a) concentration of glycerol secreted per 10⁵ adipocytes,(b) concentration of glycerol secreted per one gram of adipocytes.

FIG. 7 , which shows the results of the peptide effect confirmationexperiment according to Experimental Example 3, is images showing themeasured size of the adipocytes of a test subject for each experimentalgroup, in which Lean denotes a control group with normal weight andObese denotes an obesity group induced by high-fat diet (DAPI stainedimages, LipidTOX stained images, and Merge images are shown,respectively).

FIG. 8 , which shows the results of the peptide effect confirmationexperiment according to Experimental Example 3, is images showing themeasured size of the adipocytes of a test subject for each experimentalgroup (DAPI stained images of nuclei and lipids of adipocytes for eachexperimental group are shown), in which Lean denotes a control groupwith normal weight, Obese denotes an obesity group induced by high-fatdiet, Mock denotes a group administered with placebo, and 3H-OTP denotesGroup 2-1.

FIG. 9 , which shows the results of the peptide effect confirmationexperiment according to Experimental Example 3, is graphs showing themeasured blood lipid concentration of a test subject for eachexperimental group, in which TG denotes triglyceride, NEFA denotesnon-esterified fatty acid, CHOL denotes cholesterol, HDL denoteshigh-density lipoprotein, and LDL denotes low-density lipoprotein.

FIG. 10 , which shows the results of the peptide effect confirmationexperiment according to Experimental Example 4, is a graph showing themeasured body weight per week of age of a test subject for eachexperimental group, in which Lean denotes a control group with normalweight, Obese denotes an obesity group induced by high-fat diet, Mockdenotes a group administered with placebo, 3H-OTP 30 μg denotes Group3-2, and 3H-OTP 50 μg denotes Group 3-1. For a reference purpose, agraph relating to 3H-OTP-2W 50 μg representing Group 2-1 is also shown.

FIG. 11 , which shows the results of the peptide effect confirmationexperiment according to Experimental Example 4, is a graph showing themeasured antibody titer per week of age of a test subject for eachexperimental group, in which Lean denotes a control group with normalweight, Obese denotes an obesity group induced by high-fat diet, Mockdenotes a group administered with placebo, 3H-OTP 30 μg denotes Group3-2, and 3H-OTP 50 μg denotes Group 3-1.

FIG. 12 , which shows the results of the peptide effect confirmationexperiment according to Experimental Example 5, is a graph showing themeasured body weight per week of age of a test subject for eachexperimental group, in which Lean denotes a control group with normalweight, Obese denotes an obesity group induced by high-fat diet, and3H-OTP denotes Group 4-1.

FIG. 13 , which shows the results of the peptide effect confirmationexperiment according to Experimental Example 6, is graphs showing theincrease of body weight at 16 weeks of age compared to that at 11 weeksof age, after measurement of the body weight per week of age of a testsubject for each experimental group by age, in which Wild(+/+)-Leandenotes a control group with normal weight, Wild(+/+) denotes Group 5-1,Hetero(+/−) denotes Group 5-2, and Homo(−/−) denotes Group 5-3.

FIGS. 14 to 22 , which show the results of the peptide effectconfirmation experiment according to Experimental Example 7, and aregraphs showing the measured body weight per week of age of a testsubject in each experimental group, in which Lean denotes a controlgroup with normal weight, Obese denotes an obesity group induced byhigh-fat diet, P1 denotes Group 6-1, P2 denotes Group 6-2, P3 denotesGroup 6-3, P4 denotes Group 6-4, P5 denotes Group 6-5, P6 denotes Group6-6, P7 denotes Group 6-7, P8 denotes Group 6-8, and P9 denotes Group6-9.

FIGS. 23 to 25 , which show the results of the peptide effectconfirmation experiment according to Experimental Example 7, and aretables describing the measured antibody titers at 11 weeks of age, 16weeks of age, and 19 weeks of age of a test subject for eachexperimental group, in which Lean denotes a control group with normalweight, Obese denotes an obesity group induced by high-fat diet, P1denotes Group 6-1, P2 denotes Group 6-2, P3 denotes Group 6-3, P4denotes Group 6-4, P5 denotes Group 6-5, P6 denotes Group 6-6, P7denotes Group 6-7, P8 denotes Group 6-8, and P9 denotes Group 6-9. Inaddition, the value identified by the label in the “No” column for eachexperimental group in each table denotes the experimental results ofeach individual subject in each experimental group, “ave” denotes theoverall average, “sd” denotes the overall standard deviation, and “se”denotes the overall standard error.

DETAILED DESCRIPTION FOR CARRYING OUT THE INVENTION

Hereinafter, the presently disclosed subject matter now will bedescribed in more detail in terms of some specific embodiments andexamples with reference to the accompanying drawings. It should be notedthat the accompanying drawings encompass some, but not all embodimentsof the present disclosure. The presently disclosed subject matter may beembodied in many different forms and should not be construed as limitedto the specific embodiments set forth herein. Rather, these embodimentsare provided so that this disclosure will satisfy applicable legalrequirements. Indeed, many modifications and other embodiments of thepresent disclosure will come to the mind of one skilled in the art towhich the presently disclosed subject matter pertains. Therefore, it isto be understood that the presently disclosed subject matter is not tobe limited to the specific embodiments disclosed and that modificationsand other embodiments are intended to be included within the scope ofthe appended claims.

Definitions of General Terms

About

As used herein, the term “about” refers to a degree close to a certainquantity, and it refers to an amount, level, value, number, frequency,percent, dimension, size, amount, weight, or length that varies by tothe extent of 30%, 25%, 20%, 25%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%,or 1% with respect to a reference amount, level, value, number,frequency, percentage, dimension, size, amount, weight, or length.

Peptide

As used herein, the term “peptide” refers to a polymer of amino acids.The term peptide refers to a form in which a small number of amino acidsare linked, and is mainly used to distinguish it from a protein. Thereis no clear standard for distinguishing proteins from peptides, but asused herein, while about 200 amino acid polymers are referred to aspeptides, those more than that are referred to as proteins, unlessotherwise defined. The term “peptide” may include all other meaningsrecognized by those skilled in the art.

Subject

As used herein, the term “subject” refers to an organism that is anobject exposed to a specific substance (e.g., peptide, etc.). Thesubject may refer to an independent organism (e.g., a human, animal,etc.) or may refer to a partial constitution of the independent organism(e.g., a part of a tissue, cell, etc.). This meaning may beappropriately interpreted according to the context. In addition, theterm “subject” may further include all meanings recognized by thoseskilled in the art.

Immunotherapeutics

As used herein, the term “immunotherapeutics” is a concept distinguishedfrom general therapeutics or vaccines. The immunotherapeutics are thesame as existing vaccines in that they are injected into the body of asubject to induce a humoral immune response against theimmunotherapeutics themselves. However, immunotherapeutics differ fromexisting vaccines in that the antibodies, which are induced as a resultof the humoral immune response, have an ability being able to bind notonly to the immunotherapeutics themselves, but also to specific tissuesand cells in the body (e.g., receptors on the cell surface) or specificsubstances (e.g., peptides, lipids, proteins, and/or saccharides)produced during metabolism; thereby they can treat a specific illness ordisease, and they can be administered continuously and repeatedly.Accordingly, the immunotherapeutics generally include antigens designedto induce antibodies having an ability to bind with a specific targettissue, cell, or substance in the body. Unless otherwise defined, theterm “immunotherapeutics” is interpreted to include all antigens thatcan be appropriately used by those skilled in the art (e.g., peptides,proteins, lipids, saccharides, and/or complexes thereof, etc.) havingthe above-described functions. The term “immunotherapeutics” may be morelimitedly referred to as “humoral-immunotherapeutics”. In addition, theterm “immunotherapeutics” may include all meanings recognized by thoseskilled in the art.

Treatment or Therapeutics

As used herein, the term “treatment” collectively refers to any director indirect action or measure to eliminate, alleviate, reduce, inhibit,or improve the disease, illness, disorder, and/or symptoms of a subject,or any direct or indirect action or measure to induce the results ofpreventing the disease, illness, disorder, and/or symptoms. As usedherein, the term “therapeutics” refers to various substances (e.g.,compounds or peptides) that can exhibit the “treatment” effect whenadministered in an appropriate way to a subject. In addition, the term“treatment” or “therapeutics” may include all other meanings recognizedby those skilled in the art.

Immunogenicity

As used herein, the term “immunogenicity” collectively refers to “theproperty of acting as an antigen capable of inducing an immune response”in the dictionary. There are various methods for measuring theimmunogenicity of a specific antigen, and the methods may beappropriately adopted or designed according to the purpose. For example,the methods may include 1) a method for confirming whether IgG, IgA,and/or IgE type antibodies are generated in the body of a subject whenthe antigen is administered into the body of the subject, 2) a methodfor confirming the time when the IgG, IgA, and/or IgE type antibodiesare generated depending on the administration cycle, 3) a method forconfirming the titer of the induced antibodies to the antigen, and 4)when the mechanism of action of the induced antibodies is found, amethod for measuring the effect according to the mechanism of action,but the methods are not limited thereto. The expression “increase ofimmunogenicity” may be used interchangeably with, for example, “increaseof the effect of inducing an immune response”, “improvement of theability to induce antibodies”, and “increase of the effectiveness as animmunotherapeutic”, and it includes all of the expressions which thoseskilled in the art can properly interpret according to the context.

Mer

As used herein, the term “mer” generally refers to the number of unitsin a high molecular weight polymer. As used herein, the term “mer” isgenerally expressed as “peptide with a length of an N mer” along with anumber when expressing the length of a peptide, which refers to apeptide in which a N number of amino acids are polymerized. The unitindicated by the expression “mer” should be properly interpreted withinthe context, and it includes all other meanings that can be recognizedby those skilled in the art.

Standard Amino Acid

As used herein, the term “standard amino acid” refers to 20 amino acidssynthesized through the transcription and translation processes of genesin the body of an organism. Specifically, the standard amino acidincludes alanine (Ala, A), arginine (Arg, R), asparagine (Asn, N),aspartic acid (Asp, D), cysteine (Cys, C), glutamic acid (Glu, E),glutamine (Gln, Q), glycine (Gly, G), histidine (His, H), isoleucine(Ile, I), leucine (Leu, L), lysine (Lys K), methionine (Met, M),phenylalanine (Phe, F), proline (Pro, P), serine (Ser, S), threonine(Thr, T), tryptophan (Trp, W), tyrosine (Tyr, Y), and valine (Val, V).The standard amino acid has a corresponding DNA codon and can berepresented by a general one-letter or three-letter notation of an aminoacid. The subjects being referred to by the term standard amino acidshould be appropriately interpreted according to the context, and theyinclude all other meanings that can be recognized by those skilled inthe art.

Nonstandard Amino Acid

As used herein, the term “nonstandard amino acid” refers to an aminoacid other than the standard amino acid. The nonstandard amino acidincludes artificial and unnatural amino acids, and it includes thoseamino acids which are chemically modified through posttranslationalmodification within an organism, etc. The nonstandard amino acidincludes, for example, D-form alanine, L-cyclohexylalanine,6-aminohexanoic acid, etc. Since the nonstandard amino acid does nothave a corresponding DNA codon, it cannot be represented by a generalone-letter or three-letter notation of an amino acid, and it is writtenusing other characters and explained via additional explanation. Thesubjects being referred to by the term nonstandard amino acids should beappropriately interpreted according to the context, and they include allother meanings that can be recognized by those skilled in the art.

Description of Peptide Sequence

Unless otherwise stated, when describing the sequence of a peptide inthe present specification, single letter notation or three letternotation of an amino acid is used, and it is written in the directionfrom the N-terminus to the C-terminus. For example, when expressed asRNVP, it refers to a peptide in which arginine, asparagine, valine, andproline are sequentially linked in the direction from the N-terminus tothe C-terminus. For another example, when expressed as Thr-Leu-Lys, itrefers to a peptide in which threonine, leucine, and lysine aresequentially linked in the direction from the N-terminus to theC-terminus. In the case of amino acids that cannot be represented by theone-letter notation, other letters are used to describe these aminoacids, and will be explained via additional explanation.

When expressing a peptide as a structural formula, N- and -C may be usedto clearly indicate the N-terminus or C-terminus, and may be underlinedso as to distinguish the N-terminus and/or C-terminus. For example, whenthe structural formula of a peptide is expressed as N-B-T-A-C, “N-”written at the beginning and “-C” written at the end are symbols toclarify the N-terminus and C-terminus directions unless otherwisespecified. This refers to a peptide in which the sequences representedby B, T, and A are linked in the direction from the N-terminus to theC-terminus.

Background—Humoral Immunity

Humoral Immunity by Immunoglobulin M (IgM)

Among humoral immunities, an IgM-induced immune response is an innateimmune function which is mainly active in the primary immune response,and it occurs rapidly in the early stages of infection. IgM is mainlysecreted in the form of a pentamer, and theoretically has 10antigen-binding sites, and thus, it can bind to a large number ofantigens simultaneously. Although IgM can bind to a wide variety oftypes and forms of antigens and, the affinity and avidity of the bindingare limited by the intrinsic affinity of IgM itself. Therefore, theaffinity and avidity of IgM to an antigen are significantly lower thanthose of an antibody, such as IgG generated by the aid of helper Tcells.

Limitations of Humoral Immunity by IgM

Although IgM-induced humoral immunity plays an important role in theinitial immune response, the effect relying on humoral immunity by IgMis limited because 1) the production of IgM produced by B cells is lowcompared to those of other types of antibodies (e.g., IgG produced bydifferentiated B cells, etc.), 2) the specific binding ability of IgM toan antigen is low compared to that of IgG, and 3) the degree of asecondary immune response is weak when re-exposed to the same antigen.Therefore, from the viewpoint of designing an antigen that induces animmune response, in the case where the antigen injected into the body ofa subject induces only a humoral immunity by IgM, it is highly likelythat the desired effect will not be obtained. Therefore, it is veryimportant to design an antigen so as to induce a humoral immunity byIgG.

Humoral Immunity by Immunoglobulin G (IgG) 1—Overview

The humoral immunity that produces IgG mainly occurs in the germinalcenter of the spleen or lymph node, and it proceeds by the complexaction of B cells, helper T cells, and antigen-presenting cells (APCs).The overall process is as follows. 1) B cells recognize invadingantigens (mainly proteins or peptide antigens). 2) After theantigen-presenting cells endocytose the antigens (or fragments thereof)and cut them into smaller fragments within the cells, and present someof the fragments to MHC Class II on the surface of theantigen-presenting cells. 3) Helper T cells recognize the antigenfragments presented to the MHC Class II. 4) The helper T cells transmita differentiation signal to the B cells (antigen-recognized cells). 5)The B cells are activated and some differentiate into plasma cells toproduce an IgG antibody having a high specific binding ability to theantigens. 6) As a result of the activation of the B cells, some cellsdifferentiate into memory B cells and are stored in the body so thatthey can trigger an immune response that quickly produces an IgG antigenwhen the same antigens re-invade.

Humoral Immunity by IgG 2—Antigen-Recognized Cells

Antigen-presenting cells are a collective term for cells which arecapable of endocytosing protein fragments or peptides, cleaving theminto shorter peptide fragments, placing them on MHC Class II, andpresenting them on the surface of the antigen-presenting cells. Majorantigen-presenting cells include B cells, macrophages, dendritic cells,etc. Antigen-presenting cells transport the endocytosed antigenfragments from the site of infection to the lymph node, and present theantigen fragments to helper T cells by MHC Class II, and thereby play arole in inducing an immune response by activating helper T cells thatrecognize the same.

Humoral Immunity by IgG 3—MHC Class II

MHC Class II is a molecule expressed on the surface ofantigen-presenting cells and has a heterodimer structure consisting ofα/β chains. MHC Class II, due to its structure, can bind to a peptide ofa certain length and present the same. Antigen-presenting cells allowpeptide fragments derived from foreign antigens to bind to MHC Class IIand present them on the cell surface. HLA gene complexes (humanleukocyte antigen gene complexes) are involved in the expression of MHCClass II in humans, and among them, gene complexes such as HLA-DP, DQ,DR, etc. are known to be involved in the expression of MHC Class II cellsurface receptors on the surface of antigen-presenting cells. In humans,the HLA-DR gene is known to have various alleles according to race, andabout 12 types of HLA-DR genes are known as the most frequently foundalleles.

Humoral Immunity by IgG 4—MHC Class II Presentation by AntigenPresenting Cells

Although there is a slight difference between the literatures, thelength of the peptide presented in MHC Class II is known to be in therange of about a 17mer to about a 24mer. Therefore, antigen-presentingcells do not present the endocytosed antigen proteins or peptidefragments on MHC Class II as they are, but they undergo a process ofcleavage making them into smaller fragments of 17mers to 24mers. Theantigen fragments (proteins or peptide fragments) endocytosed by theantigen-presenting cells are present in the endosome, and the endosomeis fused with the lysosome of the antigen-presenting cells. Thereafter,the antigen fragments are cleaved into shorter peptides by various kindsof degrading enzymes present in the lysosome. Examples of the degradingenzymes include endopeptidases and exopeptidases. While endopeptidasesact to cleave the antigen fragments by acting on the peptide bond insidethe antigen fragments, exopeptidases mainly act to cleave the antigenfragments by acting on peptide bonds at both ends of the antigenfragments. When the antigen fragments are cleaved into peptides of anappropriate size through the above process, some of them are bound toMHC Class II present in the inner membrane of the lysosome. The lysosomereturns to the cell surface and fuses with the cell's plasma membrane,and thereby MHC Class II and the peptide fragments bound thereto areexposed on the surface of antigen-presenting cells. This whole processis also called “the process by which antigen-presenting cells presentantigens”.

Humoral Immunity by IgG 5—Helper T Cell

Helper T lymphocytes are also known as CD4+ cells because they expressCD4. Helper T cells express T cell receptors (TCRs) which have theability to bind to MHC Class II on the surface. The T cell receptorsgenerally form a complex with CD3. When antigens (e.g., peptidefragments) transported by antigen-presenting cells to lymph nodes arepresented through MHC Class II, helper T cells recognize the antigenfragments presented above. The T cell receptor-CD3 complex and CD4 areinvolved in this recognition process. When the helper T cellssuccessfully recognize the antigen fragments, they are activated tosecrete various cytokines or differentiate themselves. The secretedcytokines are involved in the differentiation of B cells, which will bedescribed later.

Humoral Immunity by IgG 6—B Cell Differentiation

Under the influence of the cytokines (e.g., interleukin-4 (IL-4), etc.)secreted by helper T cells, there occurs an immunoglobulin classswitching of the B cells, thereby changing the isotypes of theantibodies produced by the B cells (e.g., from IgM to IgG). In addition,some of the B cells are differentiated into memory B cells and stored soas to induce a rapid immune response when the same antigens invadeagain, and some are differentiated into plasma cells and activelyproduce IgG antibodies.

Humoral Immunity by IgG 7 —Conditions for IgG Production

For the occurrence of a humoral immunity by IgG, it is essential that 1)a specific three-dimensional structure of an antigen be recognized by Bcells, and 2) some fragments of the antigen be recognized by helper Tcells through MHC Class II. In general, although the part of an antigenrecognized by B cells and the part of an antigen recognized by helper Tcells are different from each other, and they activate an immuneresponse through pathways different from each other, it is generallyknown that the immune response occurs only when the part recognized by Bcells (B-cell epitope) and the part recognized by helper T cells (Thepitope) have at least a certain linkage. For example, the B-cellepitope and the Th epitope may be included in one molecule, form aconjugate, or have other linkages.

Limitations of Prior Art

Things to Consider in Designing Peptides that Can be Used asImmunotherapeutics

As described above, immunotherapeutics are required due to theircharacteristics that 1) they be able to stably induce an immune responsein the body of a subject, 2) they be able to minimize side effects byuniformly inducing only the intended antibody in the body of a subject;and 3) for their commercialization, they be easily synthesized and theirproduction cost be reasonable. Therefore, in designing a peptide thatcan be used as an immunotherapeutic, the following three conditionsshould be essentially considered: 1) the peptide should exhibit acertain level of immunogenicity, 2) the peptide should trigger an immuneresponse in the body of a subject that uniformly induces antigenrecognition specificity of the antibody intended in advance, the isotypethat controls physiological functions of the antibody, etc., and 3) thepeptide should be easy to synthesize in consideration of economicfeasibility.

Limitations of Prior Art

As disclosed in previously filed patent applications U.S. Ser. No.10/378,707 and PCT/KR2005/000784, and Kim et al. (2016, Anapolipoprotein B100 mimotope prevents obesity in mice, Clinical Science130, 105-116), it is known that antibodies specific to an artificiallyproduced peptide with a specific sequence can also bind to an exposedsite of the ApoB-100 protein in an LDL molecule, and thereby it canfunction as immunotherapeutics. Using such a characteristic,immunotherapeutics including the peptide were designed and are disclosedin the above patent applications, etc. However, the prior art was mainlyfocused on improving the immunogenicity of the peptide, for example, 1)preparing a long continuous identical sequence (concatemer) of thepeptide, 2) designing an immunotherapeutic by linking a helper T cellepitope (that is sufficiently long at a protein level) to theconcatemer, etc. Accordingly, conventionally designed immunotherapeuticshad limitations in that 1) various types of antibodies were induced andthe uniformity was decreased due to the presence of various epitopes(antigenic determinants), and 2) the economic feasibility was low due totheir high production cost.

Necessity for Establishment of Method for Peptide Design

As for the peptides for use as immunotherapeutics, no principle has beenestablished with regard to 1) uniform induction of only the intendedimmune response and 2) design of a peptide that is easy to synthesizeand has a low production cost. Accordingly, in the presentspecification, technical matters to be considered in the design ofpeptides for use as immunotherapeutics and methods for designing thesame will be provided.

A Peptide

Peptide Overview

The peptides provided herein include at least one peptide unit (a blockof peptide). The peptide unit includes at least one B-cell epitope, atleast one Th epitope, and an appropriate number of auxiliary parts. Inan embodiment, the peptide may include one peptide unit. In anotherembodiment, the peptide may include two or more peptide units.

Characteristic of Peptide 1—a Peptide Unit is Included

The peptide unit is a part designed 1) to exhibit immunogenicity beyonda certain level, and 2) to uniformly induce only the intended antibodyin advance. Therefore, the peptide unit provided herein has propertiessuitable for use as an immunotherapeutic.

Characteristic of Peptide 2—Relatively Short Length of Peptide Unit

Since the peptide unit is designed with a relatively short length, it iseasy to synthesize and the production cost is low. The peptide isdesigned using a peptide unit as a component thereof, and specifically,it is designed in a form in which one or more of the peptide units arelinked. When the peptide includes only a small number of the peptideunits, the overall peptide length is short, thus having the advantage ofeasy synthesis. Even when the peptide has a relatively long sequenceincluding a plurality of peptide units, the peptide unit itself is welldesigned for easy synthesis, and thus, it is possible to prepare thepeptide in such a manner by synthesizing the peptide units in paralleland then linking these peptide units. As a result, the peptide providedherein has a characteristic of being easy to synthesize, which is thecharacteristic suitable for the use of the peptide as animmunotherapeutic, in addition to the characteristics of the peptideunit described above.

Functions of Peptide

When the peptide is injected into the body of a subject, it has afunction of uniformly inducing only antibodies capable of specificallybinding to the B-cell epitope included in the peptide.

B-Cell Epitope

Definition of B-Cell Epitope

The peptides provided herein include one or more B-cell epitopes. Asused herein, the term B-cell epitope refers to a unit of peptide that isintentionally designed to induce a homogeneous antibody of one type.Therefore, when the peptide including the B-cell epitope is injectedinto the body of a subject, it results in that one type of antibody isdominantly induced per type of a B-cell epitope.

Structure of B-Cell Epitope

The B-cell epitope includes a part for forming a three-dimensionalstructure and an adjacent part thereof. The part for forming athree-dimensional structure is the part that forms a peptide with ahigher order structure, and this part is designed so that B cells canrecognize the peptide with a higher order structure and produce anantibody that can specifically bind to the same. The adjacent part is apart which directly or indirectly influences the part for forming athree-dimensional structure to stably form a higher order structure.Specifically, the adjacent part may have functions such as 1) a functionof the part for forming a three-dimensional structure to form a specificstructure, 2) a linker function that does not affect the part forforming a three-dimensional structure when the B-cell epitope is linkedto another part within a peptide unit, 3) a function of protecting thepart for forming a three-dimensional structure, etc., but theirfunctions are not limited thereto. In an embodiment, the B-cell epitopemay have a sequence in which a first part for forming athree-dimensional structure and a first adjacent part thereof is linkedin order from the N-terminus to the C-terminus. In another embodiment,the B-cell epitope may have a sequence in which a second adjacent part,a second part for forming a three-dimensional structure, and a thirdadjacent part are sequentially linked in order from the N-terminus tothe C-terminus. In still another embodiment, the B-cell epitope may havea sequence in which a third part for forming a three-dimensionalstructure and a fourth adjacent part are sequentially linked in orderfrom the N-terminus to the C-terminus.

Design of B-Cell Epitope 1—Designing of Part for FormingThree-Dimensional Structure

The B-cell epitope should be able to uniformly induce the production ofantibodies capable of recognizing the three-dimensional structure of Bcells and specifically binding thereto. The three-dimensional structurerecognized by B cells can be expressed through an appropriate peptidewith a higher order structure. Therefore, the B-cell epitope is designedto include a part for forming a three-dimensional structure that forms apeptide with a higher order structure. The part for forming athree-dimensional structure may form an intended peptide with a higherorder structure depending on the purpose. In an embodiment, the part forforming a three-dimensional structure may include an α-helix structure.In another embodiment, the part for forming a three-dimensionalstructure may include a β structure. In still another embodiment, thepart for forming a three-dimensional structure may include α-helixand/or β structures. In another embodiment, the part for forming athree-dimensional structure may include a peptide with a tertiarystructure. In still another embodiment, the part for forming athree-dimensional structure may include a peptide with a quaternarystructure.

Design of B-Cell Epitope 2—Adjacent Part

When designing the B-cell epitope, it is not essential that allsequences form a peptide with a higher order structure. In other words,the B-cell epitope may be designed to additionally include an adjacentpart, in addition to the part for forming a three-dimensional structure.The adjacent part may affect the part for forming a three-dimensionalstructure so that it stably forms a higher order structure. The adjacentpart may perform various other functions, and its role may overlap withthat of an auxiliary part. In an embodiment, the adjacent part may havea linker function. In another embodiment, the adjacent part may have aprotective function for the part for forming a three-dimensionalstructure. In still another embodiment, the adjacent part may have oneor more functions.

Length of B-Cell Epitope

The B-cell epitope should have 1) a size large enough to be recognizedby B cells and 2) one type or a very few types of antibodies thatspecifically binds to the B-cell epitope. The length of the B-cellepitope should be limited to an appropriate level. When the length ofthe B-cell epitope is too short, it is not recognized by B cells andthus does not have an antibody inducing ability, whereas when the lengthof the B-cell epitope is too long, various types of antibodies may beinduced, which deviates from the intended purpose. In an embodiment, thelength of the B-cell epitope may be about 8mer, about 9mer, about 10mer,about 11mer, about 12mer, about 13mer, about 14mer, about 15mer, about16mer, about 17mer, about 18mer, about 20mer, about 21mer, about 22mer,about 23mer, about 24mer, about 25mer, about 26mer, about 27mer, about28mer, about 29mer, or about 30mer.

In another embodiment, the length of the B-cell epitope may have a valuewithin the two numerical ranges selected in the immediately precedingsentence.

Embodiments of B-Cell Epitope

In an embodiment, the B-cell epitope may be one that induces an antibodytargeting apolipoprotein B-100. In another embodiment, the B-cellepitope may be a fragment of apolipoprotein B-100, and/or a mimotope ofapolipoprotein B-100. In still another embodiment, the B-cell epitope ischaracterized in that it induces an antibody that targets a siteselected from the following:

-   an externally exposed site of apolipoprotein B-100 included in    low-density lipoprotein (LDL); and an externally exposed site of    apolipoprotein B-100 included in very low-density lipoprotein    (VLDL).

Embodiments of Sequences of B-Cell Epitope

In an embodiment, the B-cell epitope is a peptide includes a sequenceselected from a group consisting of RNVPPIFNDVYWIAF(SEQ ID NO: 6),CRFRGLISLSQVYLS(SEQ ID NO: 7), KTTKQSFDLSVKAQYKKNKH(SEQ ID NO: 8),RNVPPIFNDVY(SEQ ID NO: 9), CRFRGLISLSQ(SEQ ID NO: 10), KTTKQSFDLSVK(SEQID NO: 11), RNVPPIFNDVYW(SEQ ID NO: 12), CRFRGLISLSQV(SEQ ID NO: 13),KTTKQSFDLSVKAQYKK(SEQ ID NO: 14), RNVPPIFNDVYWI(SEQ ID NO: 15),CRFRGLISLSQVY(SEQ ID NO: 16), KTTKQSFDLSVKAQYKKN(SEQ ID NO: 17),PIFNDVYWIAF(SEQ ID NO: 18), GLISLSQVYLS(SEQ ID NO: 19),QSFDLSVKAQYKKNKH(SEQ ID NO: 20), PPIFNDVYWIAF(SEQ ID NO: 21),RGLISLSQVYLS(SEQ ID NO: 22), KQSFDLSVKAQYKKNKH(SEQ ID NO: 23),VPPIFNDVYWIAF(SEQ ID NO: 24), FRGLISLSQVYLS(SEQ ID NO: 25),TKQSFDLSVKAQYKKNKH(SEQ ID NO: 26), NVPPIFNDVYWIA(SEQ ID NO: 27),RFRGLISLSQVYL(SEQ ID NO: 28), TKQSFDLSVKAQYKKN(SEQ ID NO: 29),VPPIFNDVYWI(SEQ ID NO: 30), FRGLISLSQVY(SEQ ID NO: 31),TKQSFDLSVKAQYKKN(SEQ ID NO: 32), PPIFNDVYW(SEQ ID NO: 33), RGLISLSQV(SEQID NO: 34), KQSFDLSVKAQYKK(SEQ ID NO: 35), RFRGLISLSQVYLDP(SEQ ID NO:221), SVCGCPVGHHDVVGL(SEQ ID NO: 222).

In another embodiment, the B-cell epitope may be a peptide whichincludes an epitope that is included in a peptide selected from thegroup consisting of SEQ ID NOS: 6 to 35 and 221 to 222.

Sequences Similar to Exemplary Sequences of B-Cell Epitope

In the present specification, sequences similar to the exemplarysequences of the B-cell epitope are disclosed. In an embodiment, theB-cell epitope may have a sequence having an identity of 80%, 81%, 82%,83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%,97%, 98%, 99%, or 100%, to those sequences selected from the groupconsisting of SEQ ID NOS: 6 to 34 and SEQ ID NO: 221 to 222. In anotherembodiment, the B-cell epitope may be a sequence that matches theselected sequence by more than a value selected in the immediatelypreceding sentence. For example, the B-cell epitope may have a sequencethat is 90% or more identical to SEQ ID NO: 6.

Th Epitope

Definition of Th Epitope

The Th epitope included in the peptide provided herein refers to a partwhich is designed such that after the peptide is endocytosed byantigen-presenting cells, it binds to MHC Class II, is presented on thesurface of the antigen-presenting cell, and functions to be recognizedby helper T cells (Th, helper-T-lymphocytes), in the process of beingpresented on the surface of the antigen-presenting cells by MHC ClassII. The process of presenting antigens in which antigen-presenting cellsprocess the endocytosed peptide and allow it to bind to MHC Class II wasdescribed previously. In other words, the Th epitope is a part thatplays a role to be recognized by helper T cells when the peptide isinjected into the body of a subject; therefore, it plays a direct rolein inducing an IgG-type antibody against the peptide.

Design of Th Epitope 1—Anchor Residue

The Th epitope is designed to have an anchor residue capable of bindingto MHC Class II in its sequence. Whether an anchor residue is includedin the sequence is an important factor that affects the function of theTh epitope. In an embodiment, the Th epitope may include as anchorresidues one or more amino acids selected from the group consisting oftyrosine (Y), phenylalanine (F), tryptophan (W), arginine (R), leucine(L), valine (V), isoleucine (I), and methionine (M).

Design of Th Epitope 2—Species-Specific Th Epitope

As the Th epitope, a Th epitope having an ability to bind to MHC ClassII of a certain species may be selected according to its purpose. In anembodiment, the Th epitope may be a Th epitope having the ability tobind to human MHC Class II. In another embodiment, the Th epitope may bea Th epitope having the ability to bind to MHC Class II of a speciesbelonging to a mammal. Specifically, the Th epitope may be a Th epitopehaving the ability to bind to MHC Class II of a mouse.

Design of Th Epitope 3—Gene-Specific Th Epitope

Due to the diverse traits of HLA gene complex, the structure of MHCClass II may vary between races and individuals. Accordingly, it ispossible to design a Th epitope having an ability to bind to HLA-DP,HLA-DQ, and/or HLA-DR, which are MHC Class II molecules of a specificgenetic trait. In an embodiment, the Th epitope may be a peptidesequence, which has a high binding ability to MHC Class II expressed byone or more HLA-DR genes selected from 2w2b, 2w2a, 3, 4w4, 4w14, 5, 7,52a, 52b, 52c, and 53, which are HLA-DR1 alleles.

In an embodiment, the Th epitope may be a peptide sequence, which has ahigh binding ability to MHC Class II expressed by one or more genesselected from HLA-DQ5, HLA-DR, HLA-DR1 to HLA-DR8, HLA-DR11, HLA-DR13,HLA-DR14, HLA-DRw52, HLA-DR2w15, HLA-DPw4, each subtype of HLA-DRB1(e.g., 0301, 01, 03, 04, 07, 08, 09, 11, 12, 13, 15, and 0301), andHLA-DRB5.

In another embodiment, the Th epitope may be a sequence named HA307-312disclosed in Cara C. Wilson et al. (2001, Identification andAntigenicity of Broadly Cross-Reactive and Conserved HumanImmunodeficiency Virus Type 1-Derived Helper T-Lymphocyte Epitopes,Journal of Virology, 75(9) 4195-4207).

In still another embodiment, the Th epitope may be one of the HLA ClassII restricted epitopes disclosed in Table 2 of Christopher P Desmond etal. (2008, A systematic review of T-cell epitopes in hepatitis B virus:identification, genotypic variation and relevance to antiviraltherapeutics, Antiviral Therapy 13:161-175).

Design of Th Epitope 4—Gene-Nonspecific Th Epitope

Irrespective of the traits of the HLA gene complex, Th epitopes havingan ability to bind to various MHC Class II are known, and it is possibleto design a Th epitope that can bind to various MHC Class II regardlessof genetic traits. In an embodiment, the Th epitope may be a sequencenamed “pan DR-binding peptide” disclosed in the U.S. patent applicationSer. No. 305,871.

Design of Th Epitope 5—Excluding Possibility Acting as B-Cell Epitope

The Th epitope is designed to be presented by MHC Class II ofantigen-presenting cells and recognized by helper T cells. Therefore,the Th epitope generally has a very high binding capacity to MHC ClassII, and thus, the probability that the Th epitope may act as a B-Cellepitope is very low. In other words, the Th epitope is designed suchthat it does not induce an antibody which specifically binds to thethree-dimensional structure of the Th epitope itself.

Design of Length of Th Epitope

The Th epitope should be designed to have an appropriate length so thatit can bind to one unit of MHC Class II. It is generally known that oneunit of Th epitope that can directly bind to MHC Class II is at a lengthof about a 30mer (Abbas, A. K., Lichtman, A. H. and Pillai, S. Cellularand molecular immunology (pp 124-126), 7th Ed. (2012) Philadelphia Pa.,Elsevier Saunder, etc.). In addition, 1) when the length of the Thepitope becomes too short, there is a risk for the Th epitope to losethe ability to bind to MHC Class II, whereas 2) when the length of theTh epitope is too long, there is a room for the Th epitope to actindependently as a B-cell epitope, thus deviating from the intendedpurpose. Therefore, it is necessary to design a Th epitope with anappropriate length.

Range of Length of Th Epitope

In an embodiment, the length of Th epitope may be 7mer, 8mer, 9mer,10mer, 11mer, 12mer, 13mer, 14mer, 15mer, 16mer, 17mer, 18mer, 19mer,20mer, 21mer, 22mer, 23mer, 24mer, 25mer, 26mer, 27mer, 28mer, 29mer,30mer, 31mer, 32mer, or 33mer. In another embodiment, the length of Thepitope may have a value within the two numerical ranges selected in theimmediately preceding sentence. For example, the length of Th epitopemay be in the range of 8mer to 32mer. For another example, the length ofTh epitope may be in the range of 11mer to 13mer.

Embodiments of Th Epitope—Design of PADRE

In an embodiment, the Th epitope may be a peptide named “pan DR-bindingpeptide” disclosed in U.S. patent application Ser. No. 305,871. Inanother embodiment, the Th epitope may be one of the peptides disclosedin Tables VIII A and IX of U.S. Pat. No. 6,413,935 B1. In still anotherembodiment, the Th epitope may have a peptide sequence satisfying thefollowing structural Formula I:

N-Lys-X₁-X₂-Ala-Ala-X₃-Thr-X₄-X₅-Ala-Ala-C  [Formula 1]

in which the X₁ may be tyrosine (Tyr), phenylalanine (Phe), orL-cyclohexylalanine, but the X₁ is not limited thereto.

The X₂ may be a hydrophobic amino acid, or may be leucine (Leu) orisoleucine (Ile), but the X₂ is not limited thereto.

The X₃ may be an aromatic or cyclic amino acid, or may be phenylalanine(Phe), tyrosine (Tyr), or histidine (His), but the X₃ is not limitedthereto.

The X₄ may be an aliphatic long chain amino acid, or may be isoleucine(Ile) or valine (Val), but the X₄ is not limited thereto.

X₅ may be a charged amino acid, or may be arginine (Arg), leucine (Leu),aspartic acid (Asp), glutamine (Gln), or glycine (Gly), but the X₅ isnot limited thereto.

In an embodiment, the Th epitope may have a peptide sequence satisfyingthe following structural Formula II:

N-X₁-X₂-Val-X₃-Ala-X₄-Thr-Leu-Lys-Ala-Ala-C  [Formula II]

in which the X₁ is lysine (Lys) or arginine (Arg),

the X₂ is tyrosine (Tyr), phenylalanine (Phe), or L-cyclohexylalanine,

the X₃ is lysine (Lys), tryptophan (Trp), tyrosine (Tyr), arginine(Arg), alanine (Ala), or methionine (Met), and

the X₄ is asparagine (Asn), tryptophan (Trp), tyrosine (Tyr), valine(Val), histidine (His), lysine (Lys), or alanine (Ala).

Embodiments of Sequences of Th Epitope

In an embodiment, the Th epitope may be selected from a group consist ofK(Cha)VAAWTLKAA(SEQ ID NO: 1), PKYVKQNTLKLAT(SEQ ID NO: 2),ILMQYIKANSKFIGI(SEQ ID NO: 3), QSIALSSLMVAQAIP(SEQ ID NO: 4),ILMQYIKANSKFIGIPMGLPQSIALSSLMVAQ(SEQ ID NO: 5), PLGFFPDHQL(SEQ ID NO:162), WPEANQVGAGAFGPGF(SEQ ID NO: 163), MQWNSTALHQALQDP(SEQ ID NO: 164),MQWNSTTFHQTLQDPRVRGLYFPAGG(SEQ ID NO: 165), FFLLTRILTI(SEQ ID NO: 166),FFLLTRILTIPQSLD(SEQ ID NO: 167), TSLNFLGGTTVCLGQ(SEQ ID NO: 168), 17QSPTSNHSPTSCPPIC(SEQ ID NO: 169), IIFLFILLLCLIFLLVLLD(SEQ ID NO: 170),CTTPAQGNSMFPSC(SEQ ID NO: 171), CTKPTDGN(SEQ ID NO: 172), WASVRFSW(SEQID NO: 173), LLPIFFCLW(SEQ ID NO: 174), MDIDPYKEFGATVELLSFLP(SEQ ID NO:175), FLPSDFFPSV(SEQ ID NO: 176), RDLLDTASALYREALESPEH(SEQ ID NO: 177),PHHTALRQAILCWGELMTLA(SEQ ID NO: 178), GRETVIEYLVSFGVW(SEQ ID NO: 179),EYLVSFGVWIRTPPA(SEQ ID NO: 180), VSFGVWIRTPPAYRPPNAPI(SEQ ID NO: 181),TVVRRRGRSP(SEQ ID NO: 182), VGPLTVNEKRRLKLI(SEQ ID NO: 183),RHYLHTLWKAGILYK(SEQ ID NO: 184), ESRLVVDFSQFSRGN(SEQ ID NO: 185),LQSLTNLLSSNLSWL(SEQ ID NO: 186), SSNLSWLSLDVSAAF(SEQ ID NO: 187),LHLYSHPIILGFRKI(SEQ ID NO: 188), KQCFRKLPVNRPIDW(SEQ ID NO: 189),LCQVFADATPTGWGL(SEQ ID NO: 190), AANWILRGTSFVYVP(SEQ ID NO: 191),EIRLKVFVLGGCRHK(SEQ ID NO: 192), KFVAAWTLKAA(SEQ ID NO: 195),KYVAAWTLKAA(SEQ ID NO: 196), DIEKKIAKMEKASSVFNVVNS(SEQ ID NO: 223),YSGPLKAEIAQRLEDV(SEQ ID NO: 224), K(Cha)VKANTLKAA(SEQ ID NO: 225),K(Cha)VKANTLKAA(SEQ ID NO: 226), K(Cha)VKAWTLKAA(SEQ ID NO: 227),K(Cha)VKAWTLKAA(SEQ ID NO: 228), K(Cha)VWANTLKAA(SEQ ID NO: 229),K(Cha)VWANTLKAA(SEQ ID NO: 230), K(Cha)VWAYTLKAA(SEQ ID NO: 231),K(Cha)VWAVTLKAA(SEQ ID NO: 232), K(Cha)VYAWTLKAA(SEQ ID NO: 233),K(Cha)VYAWTLKAA(SEQ ID NO: 234), R(Cha)VRANTLKAA(SEQ ID NO: 235),K(Cha)VKAHTLKAA(SEQ ID NO: 236), K(Cha)VKAHTLKAA(SEQ ID NO: 237),K(Cha)VAANTLKAA(SEQ ID NO: 238), K(Cha)VAANTLKAA(SEQ ID NO: 239),K(Cha)VAAYTLKAA(SEQ ID NO: 240), K(Cha)VAAYTLKAA(SEQ ID NO: 241),K(Cha)VAAWTLKAA(SEQ ID NO: 242), K(Cha)VAAKTLKAA(SEQ ID NO: 243),K(Cha)VAAHTLKAA(SEQ ID NO: 244), K(Cha)VAAATLKAA(SEQ ID NO: 245),K(Cha)VAAWTLKAA(SEQ ID NO: 246), and K(Cha)VMAATLKAA(SEQ ID NO: 247). Inthis case, “a” denotes D-form alanine, “Z” denotes 6-aminohexanoic acid,and “(Cha)” denotes L-cyclohexylalanine.

Sequences Similar to Exemplary Sequences of Th Epitope

In the present specification, sequences similar to the exemplarysequences of the Th epitope are disclosed. In an embodiment, the Thepitope may have a sequence, which have an identity of 80%, 81%, 82%,83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%,97%, 98%, 99%, or 100% to SEQ ID NO: 1 to SEQ ID NO: 5, SEQ ID NO: 162to SEQ ID NO: 192, SEQ ID NO: 195 to SEQ ID NO: 196, and SEQ ID NO: 223to SEQ ID NO: 247, a sequence satisfying the above [Formula I], or asequence satisfying the above [Formula II]. In another embodiment, theTh epitope may have a sequence which matches, by the number selected inthe immediately preceding sentence or more, to SEQ ID NO: 1 to SEQ IDNO: 5, SEQ ID NO: 162 to SEQ ID NO: 192, SEQ ID NO: 195 to SEQ ID NO:196, and SEQ ID NO: 223 to SEQ ID NO: 247, a sequence satisfying theabove [Formula I], or a sequence satisfying the above [Formula II]. Forexample, the Th epitope may have a sequence which has an identity of 90%or more to the sequence of SEQ ID NO: 1.

Auxiliary Part

Definition of Auxiliary Part

The peptides disclosed herein may include one or more auxiliary parts.The auxiliary part collectively refers to an additional part which candirectly or indirectly affect the peptide to cause an intended immuneresponse in the body of a subject. The auxiliary part may have one ormore functions, and the constitution of the peptide sequence and/or theposition of the sequence may appropriately be designed according to thepurpose.

Function of Auxiliary Part 1—Function as Linker

The auxiliary part may function as a linker linking a B-cell epitope anda Th epitope. The B-cell epitope and the Th epitope may be directlylinked or they may be linked through an auxiliary part serving as alinker. In addition, the auxiliary part may be designed to have a linkerfunction that links a plurality of units included in the peptide. In anembodiment, the sequence of the auxiliary part may be located betweenthe sequence of the B-cell epitope and the sequence of the Th epitope.In particular, the auxiliary part has a linker function that links theB-cell epitope and the Th epitope. In another embodiment, the sequenceof the auxiliary part may be located between the sequence of the firstpeptide unit and the sequence of the second peptide unit in the peptide.In particular, the auxiliary part has a linker function for linking thefirst peptide unit and the second peptide unit.

Function of Auxiliary Part 2—Function for Protection

In the case of the peptide unit provided herein, it is characterized byhaving a relatively short sequence length. Accordingly, when a peptideincluding the peptide unit is injected into the body of a subject, theTh epitope sequence may be degraded before being recognized by helper Tcells, and thus, the intended immune response may not occur. In anembodiment, the protective unit may protect the Th epitope from beingcleaved by an enzyme in the body of the subject. For example, the enzymein the body of the subject may be a peptidase. Specifically, thepeptidase may be an exopeptidase and/or an endopeptidase, but thepeptidase is not limited thereto. In another embodiment, the auxiliarypart may be linked to the N-terminus and/or C-terminus of the Thepitope. In particular, the auxiliary part has a function of protectingthe Th epitope. In still another embodiment, the auxiliary part mayinclude at least one nonstandard amino acid.

Function of Auxiliary Part 3—Function for Forming Cyclic Form

The auxiliary part may be designed to be linked to both ends of thepeptide unit to thereby have a function of allowing the peptide to forma cyclic form. In an embodiment, the peptide may include a firstauxiliary part at the N-terminus and a second auxiliary part at theC-terminus. In particular, the first auxiliary part and the secondauxiliary part may each include one or more cysteines (S). In anotherembodiment, the peptide may exist in a cyclic form. In particular, theN-terminus and C-terminus of the peptide may be linked through anauxiliary part.

Function of Auxiliary Part 3—Other Functions

The auxiliary part may have an additional function in addition to theabove functions. In an embodiment, the auxiliary part may include ahydrophilic amino acid and may have a function of increasing thesolubility of a peptide. In another embodiment, the auxiliary part mayconsist of a sequence that is biologically inactive in the body of asubject. In particular, the auxiliary part has no effect on thefunctions of the B-cell epitope and the Th epitope, and may have a dummyfunction to extend the length of the peptide. Specifically, the peptidemay be a His-tag, but is not limited thereto.

Capable of Performing Multiple Functions

The auxiliary part may have one or more functions. In an embodiment, theauxiliary part may have a linker function, a protective function, acyclic form forming function, a dummy function, and/or a solubilityincreasing function.

May Include Nonstandard Amino Acid

The auxiliary part may include one or more nonstandard amino acids. Theartificial amino acid may be necessary for an auxiliary part to exhibitthe linker function, protective function, and/or other functions. In anembodiment, the auxiliary part may include at least one nonstandardamino acid. Specifically, the nonstandard amino acid may be one or morenonstandard amino acids selected from the group consisting ofL-cyclohexylalanine, D-form alanine, and 6-aminohexanoic acid, but thenonstandard amino acid is not limited thereto.

Length of Auxiliary Part

The auxiliary part may be designed to have an appropriate lengthaccording to its function. When the auxiliary part has multiplefunctions, it may be designed to have an appropriate length to exhibitall of the multiple functions. In an embodiment, the length of theauxiliary part may be 1mer, 2mer, 3mer, 4mer, 5mer, timer, 7mer, 8mer,9mer, 10mer, 11mer, 12mer, 13mer, 14mer, 15mer, 16mer, 17mer, 18mer,19mer, 20mer, 21mer, 22mer, 23mer, 24mer, 25mer, 26mer, 27mer, 28mer,29mer, 30mer, or 31mer or longer. In another embodiment, the length ofthe auxiliary part may have a value within the two numerical ranges ofthe immediately preceding sentence. For example, the length of theauxiliary part may be in the range of 1mer to 8mer. For another example,the length of the auxiliary part may be in the range of 15mer to 26mer.

Characteristics of Auxiliary Part—Has Little Effect on Function ofB-Cell Epitope

The auxiliary part does not significantly affect the function of thepeptide unit and/or peptide disclosed herein to induce an antibody thatspecifically binds to the B-cell epitope in the body of a subject.

Embodiments of Sequences of Auxiliary Part

In an embodiment, the auxiliary part may be a peptide selected from agroup consisting of a, Z, aZ, Za, RN, AF, CR, LS, KT, KH, RF, DP, SV,GL, ZRNV(SEQ ID NO: 36), aZRN(SEQ ID NO: 37), IAFZ(SEQ ID NO: 38),AFZa(SEQ ID NO: 39), RNVP(SEQ ID NO: 40), WIAF(SEQ ID NO: 41), ZCRF(SEQID NO: 42), aZCR(SEQ ID NO: 43), YLSZ(SEQ ID NO: 44), LSZa(SEQ ID NO:45), CRFR(SEQ ID NO: 46), VYLS(SEQ ID NO: 47), ZKTT(SEQ ID NO: 48),aZKT(SEQ ID NO: 49), NKHZ(SEQ ID NO: 50), KHZa(SEQ ID NO: 51),GSHHHHHHGSDDDDK(SEQ ID NO: 52), HHHHHH(SEQ ID NO: 53),MRGSHHHHHHGSDDDDKIVD(SEQ ID NO: 54), GGGGSGGGGGGSS(SEQ ID NO: 55),RRRRRR(SEQ ID NO: 159), GSHHHHHHGSDDDDKaZ(SEQ ID NO: 193), andZaGSHHHHHHGSDDDDK(SEQ ID NO: 194). In particular, “a” denotes D-formalanine and “Z” denotes 6-aminohexanoic acid.

Designing Peptide Unit—Overall

A method for designing a possible peptide unit and a form thereof willbe described hereinbelow. Each unit may include at least one B-cellepitope and at least one Th epitope, and may include an appropriatenumber of auxiliary parts. The linking order of the B-cell epitope, theTh epitope, and the auxiliary part is exemplified for each type. Unlessotherwise specified, the design of each part included in the peptideunit basically follows the design principle described above.

Designing Unit-A

Structure of Unit-A1—Overview

As a peptide unit provided herein, a peptide unit, which can include 1)one B-cell epitope and one Th epitope and 2) one or more an auxiliarypart, is named “unit-A”. The function of the auxiliary part is notparticularly limited as long as it does not impair the functions of theB-cell epitope and the Th epitope, and is appropriately designed asnecessary.

In an embodiment, the unit-A may be one in which the first B-cellepitope and the first Th epitope are sequentially linked in thedirection from the N-terminus to the C-terminus.

Furthermore, the unit-A may further include a first auxiliary part. Whenthe unit-A includes the first auxiliary part, the sequence of the firstauxiliary part is located at the N-terminal side relative to thesequence of the first B-cell epitope within the unit-A sequence. Inparticular, the first auxiliary part may have a dummy function, asolubility improving function, a linker function, and/or a cyclicform-forming function, but the functions of the first auxiliary part arenot limited thereto.

Furthermore, the unit-A may further include a second auxiliary part.When the unit-A includes the second auxiliary part, the sequence of thesecond auxiliary part is located between the sequence of the firstB-cell epitope and the sequence of the first Th epitope within theunit-A sequence. In particular, the second auxiliary part may have adummy function, a solubility improving function, a linker function,and/or a protective function, but the functions of the second auxiliarypart are not limited thereto.

Furthermore, the unit-A may further include a third auxiliary part. Whenthe unit-A includes the third auxiliary part, the sequence of the thirdauxiliary part is located at the C-terminal side relative to thesequence of the first Th epitope within the unit-A sequence. Inparticular, the third auxiliary part may have a dummy function, asolubility improving function, a linker function, a protective functionand/or a cyclic form-forming function, but the functions of the thirdauxiliary part are not limited thereto.

In another embodiment, the unit-A may be one in which a second Thepitope and a second B-cell epitope are sequentially linked in thedirection from the N-terminus to the C-terminus.

Furthermore, the unit-A may further include a fourth auxiliary part.When the unit-A includes the fourth auxiliary part, the sequence of thefourth auxiliary part is located at the N-terminal side relative to thesequence of the second Th epitope within the unit-A sequence. Inparticular, the fourth auxiliary part may have a dummy function, asolubility improving function, a linker function, and/or a cyclicform-forming function, but the functions of the fourth auxiliary partare not limited thereto.

Furthermore, the unit-A may further include a fifth auxiliary part. Whenthe unit-A includes the fifth auxiliary part, the sequence of the fifthauxiliary part is located between the sequence of the second B-cellepitope and the sequence of the second Th epitope within the unit-Asequence. In particular, the fifth auxiliary part may have a dummyfunction, a solubility improving function, a linker function, and/or acyclic form-forming function, but the functions of the fifth auxiliarypart are not limited thereto.

Furthermore, the unit-A may further include a sixth auxiliary part. Whenthe unit-A includes the sixth auxiliary part, the sequence of the sixthauxiliary part is located at the C-terminal side relative to thesequence of the second B-cell epitope within the unit-A sequence. Inparticular, the sixth auxiliary part may have a dummy function, asolubility improving function, a linker function, and/or a cyclicform-forming function, but the functions of the sixth auxiliary part arenot limited thereto.

Structure of Unit-A 2—Formula

In an embodiment, the unit-A is a peptide represented by the following[Formula A] or [Formula A′].

N-A₁-B₁-A₂-T₁-A₃-C  [Formula A]

N-A₄-T₂-A₅-B₂-A₆-C  [Formula A′]

The B₁ and B₂ are B-cell epitopes, and they follow the design principledescribed above.

The T₁ and T₂ are Th epitopes, and they follow the design principledescribed above.

The A₁ to A₆ are auxiliary parts and they may be omitted.

In particular, the A₁ to A₆ may have a dummy function, a solubilityimproving function, a linker function, and/or a cyclic form formingfunction, but the A₁ to A₆ are not limited thereto.

Length of Unit-A

In an embodiment, the length of the unit-A may be about 16mer, about17mer, about 18mer, about 19mer, about 20mer, about 21mer, about 22mer,about 23mer, about 24mer, about 25mer, about 26mer, about 27mer, about28mer, about 29mer, about 30mer, about 31mer, about 32mer, about 33mer,about 34mer, about 35mer, about 36mer, about 37mer, about 38mer, about39mer, about 40mer, about 41mer, about 42mer, about 43mer, about 44mer,about 45mer, about 46mer, about 47mer, about 48mer, about 49mer, about50mer, about 51mer, about 52mer, about 53mer, about 54mer, about 55mer,about 56mer, about 57mer, about 58mer, about 59mer, about 60mer, about61mer, about 62mer, about 63mer, about 64mer, about 65mer, about 66mer,about 67mer, about 68mer, about 69mer, about 70mer, about 71mer, about72mer, about 73mer, about 74mer, about 75mer, about 76mer, about 77mer,about 78mer, about 79mer, about 80mer, about 81mer, about 82mer, about83mer, about 84mer, about 85mer, about 86mer, about 87mer, about 88mer,about 89mer, about 90mer, about 91mer, about 92mer, about 93mer, about94mer, about 95mer, about 96mer, about 97mer, about 98mer, about 99mer,or about 100mer. In another embodiment, the length of the unit-A mayhave a value within the two numerical ranges selected in the immediatelypreceding sentence. For example, the length of the unit-A may be in therange of about 16mer to about 30mer. For another example, the length ofthe unit-A may be in the range of about 23mer to about 60mer.

Embodiment of Unit-A—Exemplary Design

In an embodiment, the unit-A may have a sequence in which a first B-cellepitope, a first auxiliary part, and a first Th epitope are sequentiallylinked. In particular, the first auxiliary part has a linker functionand it includes one or more artificial amino acids.

In another embodiment, the unit-A may have a sequence in which a secondauxiliary part, a second B-cell epitope, a third auxiliary part, and asecond Th epitope are sequentially linked. In particular, the secondauxiliary part is His-tag, and the third auxiliary part has a linkerfunction and it includes one or more artificial amino acids.

In still another embodiment, the unit-A may have a sequence in which athird B-cell epitope, a fourth auxiliary part, a third Th epitope, and afifth auxiliary part are sequentially linked. In particular, the fourthauxiliary part has a linker function and a protective function, and thefifth auxiliary part has a protective function. The fourth auxiliarypart and the fifth auxiliary part each include one or more artificialamino acids.

In still another embodiment, the unit-A may have a sequence in which a6th an auxiliary part, a 4th B-cell epitope, a 7th auxiliary part, a 4thTh epitope, and an eighth auxiliary part are sequentially linked. Inparticular, the sixth auxiliary part is His-tag, the seventh auxiliarypart has a linker function and a protective function, and the eighthauxiliary part has a protective function. The seventh auxiliary part andthe eighth auxiliary part include one or more artificial amino acids.

Embodiments of Unit-A Sequence

In an embodiment, the Unit-A is a unit peptide selected from a groupconsisting of RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZ(SEQ ID NO: 56),ZaK(Cha)VAAWTLKAAaZRNVPPIFNDVYWIAF(SEQ ID NO: 57),CRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZ(SEQ ID NO: 58),ZaK(Cha)VAAWTLKAAaZCRFRGLISLSQVYLS(SEQ ID NO: 59),KTTKQSFDLSVKAQYKKNKHZaK(Cha)VAAWTLKAAaZ(SEQ ID NO: 60),ZaK(Cha)VAAWTLKAAaZKTTKQSFDLSVKAQYKKNKH(SEQ ID NO: 61),RNVPPIFNDVYWIAFK(Cha)VAAWTLKAA(SEQ ID NO: 62),K(Cha)VAAWTLKAARNVPPIFNDVYWIAF(SEQ ID NO: 63),RNVPPIFNDVYK(Cha)VAAWTLKAA(SEQ ID NO: 64),PIFNDVYWIAFK(Cha)VAAWTLKAA(SEQ ID NO: 65), PPIFNDVYWK(Cha)VAAWTLKAA(SEQID NO: 66), RNVPPIFNDVYWIAFK(Cha)VAAWTLKAAHHHHHH(SEQ ID NO: 67),RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZGSHHHHHHGSDDDDK(SEQ ID NO: 68),GSHHHHHHGSDDDDKZaK(Cha)VAAWTLKAAaZRNVPPIFNDVYWIAF(SEQ ID NO: 69),RNVPPIFNDVYWIAFGSHHHHHHGSDDDDKZaK(Cha)VAAWTLKAAaZ(SEQ ID NO: 70),GSHHHHHHGSDDDDKZaK(Cha)VAAWTLKAAaZCRFRGLISLSQVYLS(SEQ ID NO: 71),GSHHHHHHGSDDDDKCRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZ(SEQ ID NO: 72),CRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZGSHHHHHHGSDDDDK(SEQ ID NO: 73),GSHHHHHHGSDDDDKZaK(Cha)VAAWTLKAAaZKTTKQSFDLSVKAQYKKNK H(SEQ ID NO: 74),GSHHHHHHGSDDDDKKTTKQSFDLSVKAQYKKNKHZaK(Cha)VAAWTLKAA aZ(SEQ ID NO: 75),KTTKQSFDLSVKAQYKKNKHZaK(Cha)VAAWTLKAAaZGSHHHHHHGSDDDD K(SEQ ID NO: 76),MRGSHHHHHHGSDDDDKIVDRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZ (SEQ ID NO: 77),MRGSHHHHHHGSDDDDKIVDGSHHHHHHGSDDDDKRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZ(SEQ ID NO: 78),MRGSHHHHHHGSDDDDKIVDRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZGSHHHHHHGSDDDDK(SEQ ID NO: 79), RNVPPIFNDVYWIAFILMQYIKANSKFIGI(SEQ IDNO: 80), RNVPPIFNDVYWIAFILMQYIKANSKFIGIPMGLPQSIALSSLMVAQ(SEQ ID NO: 81),CRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZC(SEQ ID NO: 82),RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAACR(SEQ ID NO: 161),RNVPPIFNDVYWIAFXXKXVAAWTLKAAXXGSHHHHHHGSDDDDK(SEQ ID NO: 199),GSHHHHHHGSDDDDKXXKXVAAWTLKAAXXRNVPPIFNDVYWIAF(SEQ ID NO: 200),RNVPPIFNDVYWIAFXXKXVAAWTLKAAXX(SEQ ID NO: 204),RNVPPIFNDVYWIAFKXVAAWTLKAA(SEQ ID NO: 205),RNVPPIFNDVYWIAFKXVAAWTLKAAHHHHHH(SEQ ID NO: 206),RNVPPIFNDVYWIAFXXKXVAAWTLKAACR(SEQ ID NO: 208),RNVPPIFNDVYWIAFXXKFVAAWTLKAAXX(SEQ ID NO: 210),RNVPPIFNDVYWIAFXXKFVAAWTLKAACR(SEQ ID NO: 212),RNVPPIFNDVYWIAFCTKPTDGN(SEQ ID NO: 213), RNVPPIFNDVYWIAFLLPIFFCLW(SEQ IDNO: 214), RNVPPIFNDVYWIAFFLPSDFFPSV(SEQ ID NO: 215),RNVPPIFNDVYWIAFILMQYIKANSKFIGIHHHHHH(SEQ ID NO: 219), andRNVPPIFNDVYWIAFMDIDPYKEFGATVELLSFLPHHHHHH(SEQ ID NO: 220). In this case,the “a” denotes D-form alanine, the “Z” denotes 6-aminohexanoic acid,the “(Cha)” denotes L-cyclohexylalanine, and the “X” denotes anystandard amino acid.

Designing Unit-B

Structure of Unit-B 1—Overview

As a peptide unit provided herein, a peptide unit 1) which includes twoB-cell epitopes and one Th epitope, 2) in which the sequence of oneB-cell epitope of the two B-cell epitopes is located between thesequence of the other B-cell epitope the sequence of the Th epitope, and3) which may include one or more auxiliary parts, is named “unit-B”. Thefunction of the auxiliary part is not particularly limited as long as itdoes not impair the functions of the B-cell epitope and the Th epitope,and is appropriately designed as necessary.

In an embodiment, the unit-B may be one in which the first B-cellepitope, the second B-cell epitope, and the first Th epitope aresequentially linked in the direction from the N-terminus to theC-terminus.

Furthermore, the unit-B may further include a first auxiliary part. Whenthe unit-B includes the first auxiliary part, the sequence of the firstauxiliary part is located at the N-terminal side relative to thesequence of the first B-cell epitope within the unit-B sequence. Inparticular, the first auxiliary part may have a dummy function, asolubility improving function, a linker function, and/or a cyclicform-forming function, but the functions of the first auxiliary part arenot limited thereto.

Furthermore, the unit-B may further include a second auxiliary part.When the unit-B includes the second auxiliary part, the sequence of thesecond auxiliary part is located between the sequence of the firstB-cell epitope and the sequence of the second B-cell epitope within theunit-B sequence. In particular, the second auxiliary part may have adummy function, a solubility improving function, a linker function,and/or a protective function, but the functions of the second auxiliarypart are not limited thereto.

Furthermore, the unit-B may further include a third auxiliary part. Whenthe unit-B includes the third auxiliary part, the sequence of the thirdauxiliary part is located between the sequence of the second B-cellepitope and the sequence of the first Th epitope within the unit-Bsequence. In particular, the third auxiliary part may have a dummyfunction, a solubility improving function, a linker function, and/or aprotective function, but the functions of the third auxiliary part arenot limited thereto.

Furthermore, the unit-B may further include a fourth auxiliary part.When the unit-B includes the fourth auxiliary part, the sequence of thefourth auxiliary part is located at the C-terminal side relative to thesequence of the first Th epitope within the unit-B sequence. Inparticular, the fourth auxiliary part may have a dummy function, asolubility improving function, a linker function, and/or a cyclicform-forming function, but the functions of the fourth auxiliary partare not limited thereto.

In another embodiment, the unit-B may be one in which a second Thepitope, a third B-cell epitope, and a fourth B-cell epitope aresequentially linked in the direction from the N-terminus to theC-terminus.

Furthermore, the unit-B may further include a fifth auxiliary part. Whenthe unit-B includes the fifth auxiliary part, the sequence of the fifthauxiliary part is located at the N-terminal side relative to thesequence of the second Th epitope within the unit-B sequence. Inparticular, the fifth auxiliary part may have a dummy function, asolubility improving function, a linker function, and/or a cyclicform-forming function, but the functions of the fifth auxiliary part arenot limited thereto.

Furthermore, the unit-B may further include a sixth auxiliary part. Whenthe unit-B includes the sixth auxiliary part, the sequence of the sixthauxiliary part is located between the sequence of the second Th epitopeand the sequence of the third B-cell epitope within the unit-B sequence.In particular, the sixth auxiliary part may have a dummy function, asolubility improving function, a linker function, and/or a protectivefunction, but the functions of the sixth auxiliary part are not limitedthereto.

Furthermore, the unit-B may further include a seventh auxiliary part.When the unit-B includes the seventh auxiliary part, the sequence of theseventh auxiliary part is located between the sequence of the thirdB-cell epitope and the sequence of the fourth B-cell epitope within theunit-B sequence. In particular, the seventh auxiliary part may have adummy function, a solubility improving function, a linker function,and/or a protective function, but the functions of the seventh auxiliarypart are not limited thereto.

Furthermore, the unit-B may further include an eighth auxiliary part.When the unit-B includes the eighth auxiliary part, the sequence of theeighth auxiliary part is located at the C-terminal side relative to thesequence of the fourth B-cell epitope within the unit-B sequence. Inparticular, the eighth auxiliary part may have a dummy function, asolubility improving function, a linker function, and/or a cyclicform-forming function, but the functions of the eighth auxiliary partare not limited thereto.

Structure of Unit-B 2—Formula

In an embodiment, the unit-B is a peptide represented by the following[Formula B] or [Formula B′].

N-A₁-B₁-A₂-B₂-A₃-T₁-A₄-C  [Formula B]

N-A₅-T₂-A₆-B₃-A₇-B₄-A₈-C  [Formula B′]

The B₁ to B₄ are B-cell epitopes, and they follow the design principledescribed above.

The T₁ and T₂ are Th epitopes, and they follow the design principledescribed above.

The A₁ to A₈ are auxiliary parts and they may be omitted.

In particular, the A₁ to A₈ may have a dummy function, a solubilityimproving function, a linker function, and/or a cyclic form formingfunction, but the A₁ to A₈ are not limited thereto.

Length of Unit-B

In an embodiment, the length of the unit-B may be about 24mer, about25mer, about 26mer, about 27mer, about 28mer, about 29mer, about 30mer,about 31mer, about 32mer, about 33mer, about 34mer, about 35mer, about36mer, about 37mer, about 38mer, about 39mer, about 40mer, about 41mer,about 42mer, about 43mer, about 44mer, about 45mer, about 46mer, about47mer, about 48mer, about 49mer, about 50mer, about 51mer, about 52mer,about 53mer, about 54mer, about 55mer, about 56mer, about 57mer, about58mer, about 59mer, about 60mer, about 61mer, about 62mer, about 63mer,about 64mer, about 65mer, about 66mer, about 67mer, about 68mer, about69mer, about 70mer, about 71mer, about 72mer, about 73mer, about 74mer,about 75mer, about 76mer, about 77mer, about 78mer, about 79mer, about80mer, about 81mer, about 82mer, about 83mer, about 84mer, about 85mer,about 86mer, about 87mer, about 88mer, about 89mer, about 90mer, about91mer, about 92mer, about 93mer, about 94mer, about 95mer, about 96mer,about 97mer, about 98mer, about 99mer, or about 100mer. In anotherembodiment, the length of the unit-B may have a value within the twonumerical ranges selected in the immediately preceding sentence. Forexample, the length of the unit-B may be in the range of about 24mer toabout 45mer. For another example, the length of the unit-A may be in therange of about 40mer to about 80mer.

Embodiment of Unit-B—Exemplary Design

In an embodiment, the unit-B may have a sequence in which a firstauxiliary part, a first B-cell epitope, a second B-cell epitope, asecond auxiliary part, and a first Th epitope are sequentially linked.In particular, the first auxiliary part is His-tag, the second auxiliarypart and it includes one or more artificial amino acids.

In another embodiment, the unit-B may have a sequence in which a thirdB-cell epitope, a fourth B-cell epitope, a third auxiliary part, and asecond Th epitope are sequentially linked. In particular, the thirdauxiliary part has a linker function and it includes one or moreartificial amino acids.

In another embodiment, the unit-B may have a sequence in which a fourthauxiliary part, a fifth B-cell epitope, a sixth B-cell epitope, a fifthan auxiliary part, a third Th epitope, and a sixth auxiliary part aresequentially linked. In particular, the fourth auxiliary part isHis-tag, the fifth auxiliary part has a linker function and a protectivefunction and includes one or more artificial amino acids, and the sixthauxiliary part has a protective function and includes one or moreartificial amino acids.

In still another embodiment, the unit-B may have a sequence in which aseventh B-cell epitope, an eighth B-cell epitope, a seventh auxiliarypart, a fourth Th epitope, and an eighth auxiliary part are sequentiallylinked. In particular, the seventh auxiliary part has a linker function,a protective function, and includes one or more artificial amino acids,and the eighth auxiliary part has a protective function and includes oneor more artificial amino acids.

In still another embodiment, the unit-B may have a sequence in which aneighth auxiliary part, a ninth B-cell epitope, a ninth an auxiliarypart, a tenth B-cell epitope, a tenth auxiliary part, and a fifth Thepitope are linked in sequence. In particular, the eighth auxiliary partis His-tag, the ninth auxiliary part has a linker function, and thetenth auxiliary part has a linker function and includes one or moreartificial amino acids.

In still another embodiment, the unit-B may have a sequence in which aneleventh B-cell epitope, an eleventh auxiliary part, a twelfth B-cellepitope, a twelfth auxiliary part, and a sixth Th epitope aresequentially linked. In particular, the eleventh auxiliary part has alinker function, and the twelfth auxiliary part has a linker functionand includes one or more artificial amino acids.

In still another embodiment, the unit-B may have a sequence in which athirteenth B-cell epitope, a thirteenth auxiliary part, a fourteenthB-cell epitope, a fourteenth auxiliary part, a 7th Th epitope, and afifteenth auxiliary part are sequentially linked sequence. Inparticular, the thirteenth auxiliary part has a linker function, thefourteenth auxiliary part has a linker function and a protectivefunction, and includes one or more artificial amino acids, and thefifteenth auxiliary part has a protective function and includes one ormore artificial amino acids.

Embodiments of Unit-B Sequence

In an embodiment, the unit-B is a peptide unit selected from a groupconsisting of RNVPPIFNDVYWIAFRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZ(SEQ IDNO: 83), RNVPPIFNDVYWIAFCRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZ(SEQ ID NO:84), RNVPPIFNDVYWIAFKTTKQSFDLSVKAQYKKNKHZaK(Cha)VAAWTLKAAaZ (SEQ ID NO:85), CRFRGLISLSQVYLSRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZ(SEQ ID NO: 86),CRFRGLISLSQVYLSCRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZ(SEQ ID NO: 87),CRFRGLISLSQVYLSKTTKQSFDLSVKAQYKKNKHZaK(Cha)VAAWTLKAAaZ(SE Q ID NO: 88),KTTKQSFDLSVKAQYKKNKHRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZ 28 (SEQ ID NO:89), KTTKQSFDLSVKAQYKKNKHCRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZ(SE Q ID NO:90), KTTKQSFDLSVKAQYKKNKHKTTKQSFDLSVKAQYKKNKHZaK(Cha)VAAWT LKAAaZ(SEQ IDNO: 91), RNVPPIFNDVYWIAFCRFRGLISLSQVYLSK(Cha)VAAWTLKAA(SEQ ID NO: 92),PIFNDVYWIAFGLISLSQVYLSK(Cha)VAAWTLKAA(SEQ ID NO: 93),RNVPPIFNDVYCRFRGLISLSQK(Cha)VAAWTLKAA(SEQ ID NO: 94),PIFNDVYWIAFCRFRGLISLSQK(Cha)VAAWTLKAA(SEQ ID NO: 95),PPIFNDVYWRGLISLSQVK(Cha)VAAWTLKAA(SEQ ID NO: 96),RNVPPIFNDVYWIAFCRFRGLISLSQVYLSK(Cha)VAAWTLKAAHHHHHH(SEQ ID NO: 97),MRGSHHHHHHGSDDDDKIVDRNVPPIFNDVYWIAFCRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZ(SEQ ID NO: 98),MRGSHHHHHHGSDDDDKIVDRNVPPIFNDVYWIAFCRFRGLISLSQVYLSZaK(Ch a)VAAWTLKAA(SEQID NO: 99), RNVPPIFNDVYWIAFCRFRGLISLSQVYLSZaK(Cha)VAAWTLKAA(SEQ ID NO:100), MRGSHHHHHHGSDDDDKIVDRNVPPIFNDVYWIAFGGGGSGGGGGGSSRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAA(SEQ ID NO: 101),RNVPPIFNDVYWIAFGGGGSGGGGGGSSRNVPPIFNDVYWIAFZaK(Cha)VAA WTLKAA(SEQ ID NO:102), RNVPPIFNDVYWIAFGGGGSGGGGGGSSRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZ(SEQ ID NO: 103),RNVPPIFNDVYWIAFRNVPPIFNDVYWIAFILMQYIKANSKFIGI(SEQ ID NO: 104),RNVPPIFNDVYWIAFRNVPPIFNDVYWIAFILMQYIKANSKFIGIPMGLPQSIALS SLMVAQ(SEQ IDNO: 105), CRNVPPIFNDVYWIAFCRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZC(SEQ ID NO:106), and RNVPPIFNDVYWIAFCRFRGLISLSQVYLSXXK(Cha)VAAWTLKAAXX(SEQ ID NO:202). In this case, the “a” denotes D-form alanine, the “Z” denotes6-aminohexanoic acid, the “(Cha)” denotes L-cyclohexylalanine, and the“X” denotes any standard amino acid.

Designing Unit-C

Structure of Unit-C 1—Overview

As a peptide unit provided herein, a peptide unit 1) which includes twoB-cell epitopes and one Th epitope, 2) in which the sequence of the Thepitope is located between the sequence of one B-cell epitope of the twoB-cell epitopes and the sequence of the other B-cell epitope, and 3)which may include one or more auxiliary parts, is named “unit-C”. Thefunction of the auxiliary part is not particularly limited as long as itdoes not impair the functions of the B-cell epitope and the Th epitope,and is appropriately designed as necessary.

In an embodiment, the unit-C may be one in which the first B-cellepitope, the first Th epitope, and the second B-cell epitope aresequentially linked in the direction from the N-terminus to theC-terminus.

Furthermore, the unit-C may further include a first auxiliary part. Whenthe unit-C includes the first auxiliary part, the sequence of the firstauxiliary part is located at the N-terminal side relative to thesequence of the first B-cell epitope within the unit-C sequence. Inparticular, the first auxiliary part may have a dummy function, asolubility improving function, a linker function, and/or a cyclicform-forming function, but the functions of the first auxiliary part arenot limited thereto.

Furthermore, the unit-C may further include a second auxiliary part.When the unit-C includes the second auxiliary part, the sequence of thesecond auxiliary part is located between the sequence of the firstB-cell epitope and the sequence of the first Th epitope within theunit-C sequence. In particular, the second auxiliary part may have adummy function, a solubility improving function, a linker function,and/or a protective function, but the functions of the second auxiliarypart are not limited thereto.

Furthermore, the unit-C may further include a third auxiliary part. Whenthe unit-C includes the third auxiliary part, the sequence of the thirdauxiliary part is located between the sequence of the first Th epitopeand the sequence of the second B-cell epitope within the unit-Csequence. In particular, the third auxiliary part may have a dummyfunction, a solubility improving function, a linker function, and/or aprotective function, but the functions of the third auxiliary part arenot limited thereto.

Furthermore, the unit-C may further include a fourth auxiliary part.When the unit-C includes the fourth auxiliary part, the sequence of thefourth auxiliary part is located at the C-terminal side relative to thesequence of the second B-cell epitope within the unit-C sequence. Inparticular, the fourth auxiliary part may have a dummy function, asolubility improving function, a linker function, and/or a cyclicform-forming function, but the functions of the fourth auxiliary partare not limited thereto.

Structure of Unit-C 2—Formula

In an embodiment, the unit-C is a peptide represented by the following[Formula C].

N-A₁-B₁-A₂-T₁-A₃-B₂-A₄-C  [Formula C]

The B₁ and B2 are B-cell epitopes, and they follow the design principledescribed above.

The T₁ is a Th epitope, and it follows the design principle describedabove.

The A₁, A₂, A₃, and A₄ are auxiliary parts and they may be omitted.

In particular, the A₁, A₂, A₃, and A₄ may have a dummy function, asolubility improving function, a linker function, and/or a cyclic formforming function, but the A₁, A₂, A₃, and A₄ are not limited thereto.

Length of Unit-C

In an embodiment, the length of the unit-C may be about 24mer, about25mer, about 26mer, about 27mer, about 28mer, about 29mer, about 30mer,about 31mer, about 32mer, about 33mer, about 34mer, about 35mer, about36mer, about 37mer, about 38mer, about 39mer, about 40mer, about 41mer,about 42mer, about 43mer, about 44mer, about 45mer, about 46mer, about47mer, about 48mer, about 49mer, about 50mer, about 51mer, about 52mer,about 53mer, about 54mer, about 55mer, about 56mer, about 57mer, about58mer, about 59mer, about 60mer, about 61mer, about 62mer, about 63mer,about 64mer, about 65mer, about 66mer, about 67mer, about 68mer, about69mer, about 70mer, about 71mer, about 72mer, about 73mer, about 74mer,about 75mer, about 76mer, about 77mer, about 78mer, about 79mer, about80mer, about 81mer, about 82mer, about 83mer, about 84mer, about 85mer,about 86mer, about 87mer, about 88mer, about 89mer, about 90mer, about91mer, about 92mer, about 93mer, about 94mer, about 95mer, about 96mer,about 97mer, about 98mer, about 99mer, about 100mer. In anotherembodiment, the length of the unit-C may have a value within the twonumerical ranges selected in the immediately preceding sentence. Forexample, the length of the unit-C may be in the range of about 24mer toabout 45mer. For another example, the length of the unit-A may be in therange of about 40mer to about 80mer.

Embodiment of Unit-C—Exemplary Design

In an embodiment, the unit-C may have a sequence in which a first B-cellepitope, a first auxiliary part, a first Th epitope, a second auxiliarypart, and a second B-cell epitope are sequentially linked. Inparticular, the first auxiliary part and the second auxiliary part eachhave a linker function and a protective function. The first auxiliarypart and the second auxiliary part each include one or more artificialamino acids.

In another embodiment, the unit-C may have a sequence in which a thirdauxiliary part, a third B-cell epitope, a fourth auxiliary part, asecond Th epitope, a fifth auxiliary part, and a fourth B-cell epitopeare sequentially linked. In particular, the third auxiliary part isHis-tag, and the fourth auxiliary part and the fifth auxiliary part eachhave a linker function and a protective function. The fourth auxiliarypart and the fifth auxiliary part each include one or more artificialamino acids.

Embodiments of Unit-C Sequence

In an embodiment, the unit-C is a peptide unit selected from a groupconsisting of RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZRNVPPIFNDVYWIAF(SEQ IDNO: 107), RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZCRFRGLISLSQVYLS(SEQ ID NO:108), RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZKTTKQSFDLSVKAQYKKNKH (SEQ ID NO:109), CRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZRNVPPIFNDVYWIAF(SEQ ID NO: 110),CRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZCRFRGLISLSQVYLS(SEQ ID NO: 31 111),RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZKTTKQSFDLSVKAQYKKNKH (SEQ ID NO: 112),KTTKQSFDLSVKAQYKKNKHZaK(Cha)VAAWTLKAAaZRNVPPIFNDVYWIAF (SEQ ID NO: 113),KTTKQSFDLSVKAQYKKNKHZaK(Cha)VAAWTLKAAaZCRFRGLISLSQVYLS(SE Q ID NO: 114),KTTKQSFDLSVKAQYKKNKHZaK(Cha)VAAWTLKAAaZKTTKQSFDLSVKAQ YKKNKH(SEQ ID NO:115), PIFNDVYWIAFK(Cha)VAAWTLKAACRFRGLISLSQ(SEQ ID NO: 116),PPIFNDVYWK(Cha)VAAWTLKAARGLISLSQV(SEQ ID NO: 117), MRGSHHHHHHGSDDDDKIVDRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZRNVPPIFNDVYWIAF(SEQ ID NO: 118),MRGSHHHHHHGSDDDDKIVDRNVPPIFNDVYWIAF GGGGSGGGGGGSSILMQYIKANSKFIGIPMGLPQSIALSSLMVAQ GGGGSGGGGGGSSCRFRGLISLSQVYLS(SEQ ID NO:119), RNVPPIFNDVYWIAFILMQYIKANSKFIGICRFRGLISLSQVYLS(SEQ ID NO: 120),RNVPPIFNDVYWIAFZPKYVKQNTLKLATZCRFRGLISLSQVYLS(SEQ ID NO: 121),CRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZRNVPPIFNDVYWIAFC(SEQ ID NO: 122),RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAACRFRGLISLSQVYLS(SEQ ID NO: 160),RNVPPIFNDVYWIAFXXKXVAAWTLKAAXXCRFRGLISLSQVYLS(SEQ ID NO: 198),KTTKQSFDLSVKAQYKKNKHXXKXVAAWTLKAAXXCRFRGLISLSQVYLS(SEQ ID NO: 201),RNVPPIFNDVYWIAFXPKYVKQNTLKLATXCRFRGLISLSQVYLS(SEQ ID NO: 203),RNVPPIFNDVYWIAFXXKXVAAWTLKAACRFRGLISLSQVYLS(SEQ ID NO: 207),RNVPPIFNDVYWIAFXXKFVAAWTLKAAXXCRFRGLISLSQVYLS(SEQ ID NO: 209),RNVPPIFNDVYWIAFXXKFVAAWTLKAACRFRGLISLSQVYLS(SEQ ID NO: 211),KTTKQSFDLSVKAQYKKNKHZaWPEANQVGAGAFGPGFaZCRFRGLISLSQVY LS(SEQ ID NO:216), KTTKQSFDLSVKAQYKKNKHZaMDIDPYKEFGATVELLSFLPaZCRFRGLISLS QVYLS(SEQID NO: 217),

KTTKQSFDLSVKAQYKKNKHZaILMQYIKANSKFIGIPMGLPQSIALSSLMVAQ 32aZCRFRGLISLSQVYLS(SEQ ID NO: 218). In this case, the “a” denotes D-formalanine, the “Z” denotes 6-aminohexanoic acid, the “(Cha)” denotesL-cyclohexylalanine, and the “X” denotes any standard amino acid.

Designing Unit-D

Structure of Unit-D 1—Overview

As a peptide unit provided herein, a peptide unit, 1) which includes oneB-cell epitope and two Th epitopes, 2) in which, the sequence of one Thepitope of the two Th epitopes is located between the sequence of theother Th epitope and the sequence of the B-cell epitope, and 3) whichmay include one or more auxiliary parts, is named “unit-C”. The functionof the auxiliary part is not particularly limited as long as it does notimpair the functions of the B-cell epitope and the Th epitope, and isappropriately designed as necessary.

In an embodiment, the unit-D may be one in which a first B-cell epitope,a first Th epitope, and a second Th epitope are sequentially linked inthe direction from the N-terminus to the C-terminus.

Furthermore, the unit-D may further include a first auxiliary part. Whenthe unit-D includes the first auxiliary part, the sequence of the firstauxiliary part is located at the N-terminal side relative to thesequence of the first B-cell epitope within the unit-D sequence. Inparticular, the first auxiliary part may have a dummy function, asolubility improving function, a linker function, and/or a cyclicform-forming function, but the functions of the first auxiliary part arenot limited thereto.

Furthermore, the unit-D may further include a second auxiliary part.When the unit-D includes the second auxiliary part, the sequence of thesecond auxiliary part is located between the sequence of the firstB-cell epitope and the sequence of the first Th epitope within theunit-D sequence. In particular, the second auxiliary part may have adummy function, a solubility improving function, a linker function,and/or a protective function, but the functions of the second auxiliarypart are not limited thereto.

Furthermore, the unit-D may further include a third auxiliary part. Whenthe unit-D includes the third auxiliary part, the sequence of the thirdauxiliary part is located between the sequence of the first Th epitopeand the sequence of the second Th epitope within the unit-D sequence. Inparticular, the third auxiliary part may have a dummy function, asolubility improving function, a linker function, and/or a protectivefunction, but the functions of the third auxiliary part are not limitedthereto.

Furthermore, the unit-D may further include a fourth auxiliary part.When the unit-D includes the fourth auxiliary part, the sequence of thefourth auxiliary part is located at the C-terminal side relative to thesequence of the second Th epitope within the unit-D sequence. Inparticular, the fourth auxiliary part may have a dummy function, asolubility improving function, a linker function, and/or a cyclicform-forming function, but the functions of the fourth auxiliary partare not limited thereto.

In another embodiment, the unit-D may be one in which a third Thepitope, a fourth Th epitope, and a second B-cell epitope aresequentially linked in the direction from the N-terminus to theC-terminus.

Furthermore, the unit-D may further include a fifth auxiliary part. Whenthe unit-D includes the fifth auxiliary part, the sequence of the fifthauxiliary part is located at the N-terminal side relative to thesequence of the third Th epitope within the unit-D sequence. Inparticular, the fifth auxiliary part may have a dummy function, asolubility improving function, a linker function, and/or a cyclicform-forming function, but the functions of the fifth auxiliary part arenot limited thereto.

Furthermore, the unit-D may further include a sixth auxiliary part. Whenthe unit-D includes the sixth auxiliary part, the sequence of the sixthauxiliary part is located between the sequence of the third Th epitopeand the sequence of the fourth Th epitope within the unit-D sequence. Inparticular, the sixth auxiliary part may have a dummy function, asolubility improving function, a linker function, and/or a protectivefunction, but the functions of the sixth auxiliary part are not limitedthereto.

Furthermore, the unit-D may further include a seventh auxiliary part.When the unit-D includes the seventh auxiliary part, the sequence of theseventh auxiliary part is located between the sequence of the fourth Thepitope and the sequence of the second B-cell epitope within the unit-Dsequence. In particular, the seventh auxiliary part may have a dummyfunction, a solubility improving function, a linker function, and/or aprotective function, but the functions of the seventh auxiliary part arenot limited thereto.

Furthermore, the unit-D may further include an eighth auxiliary part.When the unit-D includes the eighth auxiliary part, the sequence of theeighth auxiliary part is located at the C-terminal side relative to thesequence of the second B-cell epitope within the unit-D sequence. Inparticular, the eighth auxiliary part may have a dummy function, asolubility improving function, a linker function, and/or a cyclicform-forming function, but the functions of the eighth auxiliary partare not limited thereto.

Structure of Unit-D 2—Formula

In an embodiment, the unit-B is a peptide represented by the following[Formula D] or [Formula D′].

N-A₁-B₁-A₂-T₁-A₃-T₂-A₄-C  [Formula D]

N-A₅-T₃-A₆-T₄-A₇-B₂-A₈-C  [Formula D′]

The B₁ and B₂ are B-cell epitopes, and they follow the design principledescribed above.

The T₁ to T₄ are Th epitopes, and they follow the design principledescribed above.

The A₁ to A₈ are auxiliary parts and they may be omitted.

In particular, the A₁ to A₈ may have a dummy function, a solubilityimproving function, a linker function, and/or a cyclic form formingfunction, but the A₁ to A₈ are not limited thereto.

Length of Unit-D

In an embodiment, the length of the unit-D may be about 24mer, about25mer, about 26mer, about 27mer, about 28mer, about 29mer, about 30mer,about 31mer, about 32mer, about 33mer, about 34mer, about 35mer, about36mer, about 37mer, about 38mer, about 39mer, about 40mer, about 41mer,about 42mer, about 43mer, about 44mer, about 45mer, about 46mer, about47mer, about 48mer, about 49mer, about 50mer, about 51mer, about 52mer,about 53mer, about 54mer, about 55mer, about 56mer, about 57mer, about58mer, about 59mer, about 60mer, about 61mer, about 62mer, about 63mer,about 64mer, about 65mer, about 66mer, about 67mer, about 68mer, about69mer, about 70mer, about 71mer, about 72mer, about 73mer, about 74mer,about 75mer, about 76mer, about 77mer, about 78mer, about 79mer, about80mer, about 81mer, about 82mer, about 83mer, about 84mer, about 85mer,about 86mer, about 87mer, about 88mer, about 89mer, about 90mer, about91mer, about 92mer, about 93mer, about 94mer, about 95mer, about 96mer,about 97mer, about 98mer, about 99mer, or about 100mer. In anotherembodiment, the length of the unit-B may have a value within the twonumerical ranges selected in the immediately preceding sentence. Forexample, the length of the unit-D may be in the range of about 24mer toabout 45mer. For another example, the length of the unit-A may be in therange of about 40mer to about 80mer.

Embodiment of Unit-D—Exemplary Design

In an embodiment, the unit-D may have a sequence in which a firstauxiliary part, a first B-cell epitope, a second auxiliary part, a firstTh epitope, a third auxiliary part, and a second Th epitope aresequentially linked. In particular, the first auxiliary part is His-tag,the second auxiliary part and the third auxiliary part each have alinker function. The second auxiliary part and the third auxiliary parteach include one or more artificial amino acids.

In another embodiment, the unit-D may have a sequence in which a fourthauxiliary part, a second B-cell epitope, a fifth auxiliary part, a thirdTh epitope, a sixth auxiliary part, a fourth Th epitope, and a seventhauxiliary part are sequentially linked. In particular, the fourthauxiliary part is His-tag, the fifth auxiliary part and the sixthauxiliary part each have a linker function, and the seventh auxiliarypart has a protective function. The fifth auxiliary part, the sixthauxiliary part, and the seventh auxiliary part each include one or moreartificial amino acids.

In still another embodiment, the unit-D includes a third B-cell epitope,an eighth auxiliary part, a fifth Th epitope, a ninth auxiliary part,and a sixth Th epitope. In particular, the eighth auxiliary part and theninth auxiliary part each have a linker function. The eighth auxiliarypart and the ninth auxiliary part each have a protective function eachinclude one or more artificial amino acids.

In still another embodiment, the unit-D includes a fourth B-cellepitope, a tenth auxiliary part, a seventh Th epitope, an eleventhauxiliary part, an eighth Th epitope, and a twelfth auxiliary part. Inparticular, the tenth auxiliary part and the eleventh auxiliary parteach have a linker function. The twelfth auxiliary part has a protectivefunction. The tenth auxiliary part, the eleventh auxiliary part, and thetwelfth auxiliary part each include one or more artificial amino acids.

Embodiments of Unit-D Sequence

In an embodiment, the unit-D is a unit peptide selected from a groupconsisting of RNVPPIFNDVYWIAF ZaK(Cha)VAAWTLKAAaZ ILMQYIKANSKFIGI(SEQ IDNO: 123), CRFRGLISLSQVYLS ZaK(Cha)VAAWTLKAAaZ ILMQYIKANSKFIGI(SEQ ID NO:124), KTTKQSFDLSVKAQYKKNKH ZaK(Cha)VAAWTLKAAaZ ILMQYIKANSKFIGI(SEQ IDNO: 125), ILMQYIKANSKFIGI ZaK(Cha)VAAWTLKAAaZ RNVPPIFNDVYWIAF(SEQ ID NO:126), ILMQYIKANSKFIGI ZaK(Cha)VAAWTLKAAaZ CRFRGLISLSQVYLS(SEQ ID NO:127), ILMQYIKANSKFIGI ZaK(Cha)VAAWTLKAAaZ KTTKQSFDLSVKAQYKKNKH(SEQ IDNO: 128), PIFNDVYWIAF K(Cha)VAAWTLKAA K(Cha)VAAWTLKAA(SEQ ID NO: 129),PPIFNDVYW K(Cha)VAAWTLKAA K(Cha)VAAWTLKAA(SEQ ID NO: 130),MRGSHHHHHHGSDDDDKIVDRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGI(SEQ ID NO: 131),MRGSHHHHHHGSDDDDKIVDILMQYIKANSKFIGIZaK(Cha)VAAWTLKAAaZ(SE Q ID NO: 132),MRGSHHHHHHGSDDDDKIVDRNVPPIFNDVYWIAFGGGGSGGGGGGSSZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGI(SEQ ID NO: 133),MRGSHHHHHHGSDDDDKIVDRNVPPIFNDVYWIAFGGGGSGGGGGGS SILMQYIKANSKFIGIPMGLPQSIALSSLMVAQGGGGSGGGGGGSSILMQYIKANSKFIGIPMGLPQSIALSSLMVAQ(SEQ ID NO: 134),RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZK(Cha)LAAFTIRAAaZ(SEQ ID NO: 135),

CRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGIC(SEQ ID NO: 136). Inthis case, the “a” denotes D-form alanine, the “Z” denotes6-aminohexanoic acid, the “(Cha)” denotes L-cyclohexylalanine, and the“X” denotes any standard amino acid.

Designing Unit-E

Structure of Unit-E1—Overview

As a peptide unit provided herein, a peptide unit 1) which includes twoB-cell epitopes and two Th epitopes, 2) in which each sequence of thetwo Th epitopes is located between the sequence of one B-cell epitope ofthe two B-cell epitopes and the sequence of the other B-cell epitope,and 3) which may include one or more auxiliary parts, is named “unit-E”.The function of the auxiliary part is not particularly limited as longas it does not impair the functions of the B-cell epitope and the Thepitope, and is appropriately designed as necessary.

In an embodiment, the unit-E may be one in which a first B-cell epitope,a first Th epitope, a second Th epitope, and a second B-cell epitope aresequentially linked in the direction from the N-terminus to theC-terminus.

Furthermore, the unit-E may further include a first auxiliary part. Whenthe unit-E includes the first auxiliary part, the sequence of the firstauxiliary part is located at the N-terminal side relative to thesequence of the first B-cell epitope within the unit-E sequence. Inparticular, the first auxiliary part may have a dummy function, asolubility improving function, a linker function, and/or a cyclicform-forming function, but the functions of the first auxiliary part arenot limited thereto.

Furthermore, the unit-E may further include a second auxiliary part.When the unit-E includes the second auxiliary part, the sequence of thesecond auxiliary part is located between the sequence of the firstB-cell epitope and the sequence of the first Th epitope within theunit-E sequence. In particular, the second auxiliary part may have adummy function, a solubility improving function, a linker function,and/or a protective function, but the functions of the second auxiliarypart are not limited thereto.

Furthermore, the unit-E may further include a third auxiliary part. Whenthe unit-E includes the third auxiliary part, the sequence of the thirdauxiliary part is located between the sequence of the first Th epitopeand the sequence of the second Th epitope within the unit-E sequence. Inparticular, the third auxiliary part may have a dummy function, asolubility improving function, a linker function, and/or a protectivefunction, but the functions of the third auxiliary part are not limitedthereto.

Furthermore, the unit-E may further include a fourth auxiliary part.When the unit-E includes the fourth auxiliary part, the sequence of thefourth auxiliary part is located between the sequence of the secondB-cell epitope and the second B-cell epitope within the unit-E sequence.In particular, the fourth auxiliary part may have a dummy function, asolubility improving function, a linker function, and/or a protectivefunction, but the functions of the fourth auxiliary part are not limitedthereto.

Furthermore, the unit-E may further include a fifth auxiliary part. Whenthe unit-E includes the fifth auxiliary part, the sequence of the fifthauxiliary part is located at the N-terminal side relative to thesequence of the second B-cell epitope within the unit-E sequence. Inparticular, the fifth auxiliary part may have a dummy function, asolubility improving function, a linker function, and/or a cyclicform-forming function, but the functions of the fifth auxiliary part arenot limited thereto.

Structure of Unit-E 2—Formula

In an embodiment, the unit-E is a peptide represented by the following[Formula E].

N-A₁-B₁-A₂-T₁-A₃-T₂-A₄-B₂-A₅-C  [Formula E]

The B₁ and B₂ are B-cell epitopes, and they follow the design principledescribed above.

The T₁ and T₂ are Th epitopes, and they follow the design principledescribed above.

The A₁, A₂, A₃, A₄, and A₅ are auxiliary parts and they may be omitted.

In particular, the A₁, A₂, A₃, A₄, and A₅ may have a dummy function, asolubility improving function, a linker function, and/or a cyclic formforming function, but the A₁, A₂, A₃, A₄, and A₅ are not limitedthereto.

Length of Unit-E

In an embodiment, the length of the unit-E may be about 32mer, about33mer, about 34mer, about 35mer, about 36mer, about 37mer, about 38mer,about 39mer, about 40mer, about 41mer, about 42mer, about 43mer, about44mer, about 45mer, about 46mer, about 47mer, about 48mer, about 49mer,about 50mer, about 51mer, about 52mer, about 53mer, about 54mer, about55mer, about 56mer, about 57mer, about 58mer, about 59mer, about 60mer,about 61mer, about 62mer, about 63mer, about 64mer, about 65mer, about66mer, about 67mer, about 68mer, about 69mer, about 70mer, about 71mer,about 72mer, about 73mer, about 74mer, about 75mer, about 76mer, about77mer, about 78mer, about 79mer, about 80mer, about 81mer, about 82mer,about 83mer, about 84mer, about 85mer, about 86mer, about 87mer, about88mer, about 89mer, about 90mer, about 91mer, about 92mer, about 93mer,about 94mer, about 95mer, about 96mer, about 97mer, about 98mer, about99mer, or about 100mer. In another embodiment, the length of the unit-Emay have a value within the two numerical ranges selected in theimmediately preceding sentence. For example, the length of the unit-Emay be in the range of about 32mer to about 60mer. For another example,the length of the unit-A may be in the range of about 50mer to about100mer.

Embodiment of Unit-E—Exemplary Design

In an embodiment, the unit-E may have a sequence in which a firstauxiliary part, a first B-cell epitope, a second auxiliary part, a firstTh epitope, a third auxiliary part, a second Th epitope, a fourthauxiliary part, and a second B-cell epitope are sequentially linked. Inparticular, the first auxiliary part is His-tag. The second auxiliarypart and the fourth auxiliary part each have a linker function and aprotective function. The third auxiliary part has a linker function. Thesecond auxiliary part, the third auxiliary part, and the fourthauxiliary part include one or more artificial amino acids.

In another embodiment, the unit-E may have a sequence in which a thirdB-cell epitope, a fifth auxiliary part, a third Th epitope, a sixthauxiliary part, a fourth Th epitope, a seventh auxiliary part, and afourth B-cell epitope are sequentially linked. In particular, the fifthauxiliary part and the seventh auxiliary part each have a linkerfunction and a protective function. The sixth auxiliary part has alinker function. The fifth auxiliary part, the sixth auxiliary part, andthe seventh auxiliary part include one or more artificial amino acids.

Embodiments of Unit-E Sequence

In an embodiment, the unit-E is a peptide unit selected from a groupconsisting of RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGIRNVPPIFNDVYWIAF(SEQ ID NO: 137),RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGICRFRGLIS LSQVYLS(SEQ IDNO: 138), RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGIKTTKQSFDLSVKAQYKKNKH(SEQ ID NO: 139),CRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGIRNVPPIFN DVYWIAF(SEQ IDNO: 140), CRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGICRFRGLISLSQVYLS(SEQ ID NO: 141),CRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGIKTTKQSFDLSVKAQYKKNKH(SEQ ID NO: 142),KTTKQSFDLSVKAQYKKNKHZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGIRNVPPIFNDVYWIAF(SEQ ID NO: 143),KTTKQSFDLSVKAQYKKNKHZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGICRFRGLISLSQVYLS(SEQ ID NO: 144),KTTKQSFDLSVKAQYKKNKHZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGIKTTKQSFDLSVKAQYKKNKH(SEQ ID NO: 145),MRGSHHHHHHGSDDDDKIVDRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGIRNVPPIFNDVYWIAF(SEQ ID NO: 146),MRGSHHHHHHGSDDDDKIVDRNVPPIFNDVYWIAFGGGGSGGGGGGS SILMQYIKANSKFIGIPMGLPQSIALS SLMVAQILMQYIKANSKFIGIPMGLPQSIALS SLMVAQGGGGSGGGGGGS SCRFRGLISLSQVYLS(SEQ ID NO: 147),PIFNDVYWIAFK(Cha)VAAWTLKAAK(Cha)VAAWTLKAACRFRGLISLSQ(SEQ ID NO: 148),PPIFNDVYWK(Cha)VAAWTLKAAK(Cha)VAAWTLKAARGLISLSQV(SEQ ID NO: 149), andCRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGIRNVPPIF NDVYWIAFC(SEQID NO: 150). In this case, the “a” denotes D-form alanine, the “Z”denotes 6-aminohexanoic acid, the “(Cha)” denotes L-cyclohexylalanine.

Designing a Peptide

Designing a Peptide—Overview

The peptides provided herein can be designed using one or more of thepeptide units disclosed above. These peptides include one or more ofpeptide units and may include one or more types of peptide units. Forexample, the peptide may be designed by 1) including only one peptideunit, 2) designing a concatemer by linking multiple peptide units of onetype having the same sequence, 3) designing the peptide in the form ofstring-of-beads by linking one or more types of the peptide units withdifferent sequences, 4) by mixing the design methods of 1) to 3) above,and 5) designing the peptide in a cyclic form by connecting both ends ofthe peptide designed in the above method, but the design methods are notlimited thereto. Hereinafter, each design method will be described indetail.

One Unit Design

The peptide may be designed to include only one of the peptide unitsdescribed above. In an embodiment, the peptide may include one peptideunit selected from the group consisting of unit-A, unit-B, unit-C,unit-D, and unit-E. In particular, the peptide unit has the constitutiondescribed above.

Concatemer Design 1—Overview

The peptide may be designed in the form of a concatemer in whichmultiple peptide units having the same sequence are linked. The peptidedesigned in the concatemer form consists of 1) one type of a peptideunit and 2) multiple peptide units having the same or equivalentsequence.

In particular, two peptide units with “an equivalent sequence” refers tocases where 1) when an auxiliary part is present at the N-terminusand/or C-terminus of each of the two peptides, and 2) when the auxiliarypart exists, its sequence, even if the two are different, the rest ofthe sequence is the same. For example, when a first peptide has asequence in which a first auxiliary part and a first unit-A are linkedin the direction from the N-terminus to C-terminus, a second peptide hasa sequence in which the first unit-A and a first auxiliary part arelinked in the direction from the N-terminus to the C-terminus, a thirdpeptide has a sequence in which a second auxiliary part, the firstunit-A, and a third auxiliary part are linked in the direction from theN-terminus to the C-terminus, and a fourth peptide has the first unit-Asequence, the first to fourth peptides are said to have an equivalentsequence.

In an embodiment, the peptide may include one in which a first a peptideunit and a second a peptide unit are linked in order. In particular, thefirst peptide unit is a peptide unit selected from the group consistingof unit-A, unit-B, unit-C, unit-D, and unit-E; and the second a peptideunit has a sequence which is the same as or equivalent to the firstpeptide unit.

In another embodiment, the peptide may include one in which a thirdpeptide unit, a fourth peptide unit, and a fifth peptide unit are linkedin order. In particular, the third peptide unit is a peptide unitselected from the group consisting of unit-A, unit-B, unit-C, unit-D,and unit-E; and the fourth peptide unit and the fifth peptide unit eachhave a sequence which is the same as or equivalent to the third peptideunit.

In still another embodiment, the peptide may include one in which asixth peptide unit, a seventh peptide unit, an eighth peptide unit, anda ninth peptide unit are linked in order. In particular, the thirdpeptide unit is a peptide unit selected from the group consisting ofunit-A, unit-B, unit-C, unit-D, and unit-E; and the seventh peptideunit, the eighth peptide unit, and the ninth peptide unit each have asequence which is the same as or equivalent to the sixth peptide unit.

Concatemer Design 2—Formula

In an embodiment, the peptide may be a peptide represented by thefollowing [Formula 1].

N-U₁-U₂- . . . -U_(n)-C  [Formula 1]

in which U₁ to U_(n) are each a peptide unit selected from the groupconsisting of unit-A, unit-B, unit-C, unit-D, and unit-E, and they havethe constitutions of peptide units described above.

The U₁ to U_(n) have the same or equivalent sequence.

The n is an integer of 2 or greater.

Concatemer Design 3—Exemplary Sequences

In an embodiment, the peptide is a peptide selected from a groupconsisting of MRGSHHHHHHGSDDDDKIVDRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZ(SEQ ID NO: 151),RNVPPIFNDVYWIAFCRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZRNVPPIFNDVYWIAFCRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZ(SEQ ID NO: 152),RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZCRFRGLISLSQVYLSRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZCRFRGLISLSQVYLS(SEQ ID NO: 153),RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGIRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGI(SEQ ID NO: 154), andRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGIRNVPPIFNDVYWIAFRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGIRNVPPIFNDVYWIAF(SEQ ID NO: 155). In this case, the “a” denotes D-formalanine, the “Z” denotes 6-aminohexanoic acid, the “(Cha)” denotesL-cyclohexylalanine.

String-of-Beads Design 1—Overview

The peptide may be designed in the form of string-of-beads in whichmultiple peptide units having different sequences are linked. Thepeptide designed in the form of string-of-beads consists of 1) at leastone kind of a peptide unit and 2) multiple peptide units havingdifferent sequences.

In an embodiment, the peptide may include one in which the first peptideunit and the second peptide unit are sequentially linked. In particular,the first peptide unit and the second peptide unit are each a peptideunit selected from the group consisting of unit-A, unit-B, unit-C,unit-D, and unit-E; and the first peptide unit and the second peptideunit have sequences different from each other.

In another embodiment, the peptide may include one in which the thirdpeptide unit, the fourth peptide unit, and the fifth peptide unit aresequentially linked. In particular, the third peptide unit, the fourthpeptide unit, and the fifth peptide are each a peptide unit selectedfrom the group consisting of unit-A, unit-B, unit-C, unit-D, and unit-E;and the third peptide unit, the fourth peptide unit, and the fifthpeptide unit have sequences different from one other.

In still another embodiment, the peptide may include one in which thesixth peptide unit, the seventh peptide unit, the eighth peptide unit,and the ninth peptide unit are sequentially linked. In particular, thesixth peptide unit, the seventh peptide unit, the eighth peptide unit,and the ninth peptide unit are each a peptide unit selected from thegroup consisting of unit-A, unit-B, unit-C, unit-D, and unit-E; and thesixth peptide unit, the seventh peptide unit, the eighth peptide unit,and the ninth peptide unit have sequences different from one other.

String-of-Beads Design 2—Formula

In an embodiment, the peptide may be a peptide represented by thefollowing [Formula 2]:

N-U₁-U₂- . . . -U_(n)-C  [Formula 2]

in which U₁ to U_(n) are each a peptide unit selected from the groupconsisting of unit-A, unit-B, unit-C, unit-D, and unit-E, and they havethe constitutions of peptide units described above.

The U₁ to U_(n) have the same or equivalent sequence.

The n is an integer of 2 or greater.

String-of-Beads Design 3—Exemplary Sequences

In an embodiment, the peptide may be a peptide ofRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZRNVPPIFNDVYWIAFCRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZ (SEQ ID NO: 156), in which “a” denotesD-form alanine, Z denotes 6-aminohexanoic acid, and (Cha) denotesL-cyclohexylalanine.

Mixed Design 1—Overview

The peptide may be designed by appropriately mixing theabove-mentioned 1) one unit design, 2) concatemer design, and 3)string-of-beads design. In an embodiment, the peptide may be designed byfirst designing a unit peptide according to the design methods describedabove, and then linking multiple unit peptides.

In an embodiment, the peptide may be one in which a first unit peptideand a second unit peptide are sequentially linked. In particular, thefirst unit peptide and the second unit peptide each have a peptideconstitution according to any one of the one unit design, the concatemerdesign, and the string-of-beads design, and the sequence of the firstunit peptide and the sequence of the second unit peptide are differentfrom each other.

In another embodiment, the peptide may be one in which a third unitpeptide, a fourth unit peptide, and a fifth unit peptide aresequentially linked. In particular, the third unit peptide, the fourthunit peptide, and the fifth unit peptide each have a peptideconstitution according to any one of the one unit design, the concatemerdesign, and the string-of-beads design, and the sequences of the thirdunit peptide, the fourth unit peptide, and the fifth unit peptide aredifferent from one other.

In still another embodiment, the peptide may be one in which a sixthunit peptide, a seventh unit peptide, an eighth unit peptide, and aninth unit peptide are sequentially linked. In particular, the sixthunit peptide, the seventh unit peptide, the eighth unit peptide, and theninth unit peptide each have a peptide constitution according to any oneof the one unit design, the concatemer design, and the string-of-beadsdesign, and the sequences of the sixth unit peptide, the seventh unitpeptide, the eighth unit peptide, and the ninth unit peptide aredifferent from one other.

Mixed Design 2—Formula

In an embodiment, the peptide may be a peptide represented by thefollowing [Formula 3]:

N-P₁-P₂- . . . -P_(n)-C  [Formula 3]

in which P₁ to P_(n) are each a unit peptide designed by a methodselected from the group consisting of a one unit design, a concatemerdesign, and a string-of-beads design, and they have the designs andconstitutions of peptides described above.

The n is an integer of 2 or greater.

Mixed Design 3—Exemplary Sequences

In an embodiment, the peptide is a peptide selected from a groupconsisting of RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZRNVPPIFNDVYWIAFZRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGIZRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGICRFRGLISLSQVYLS(SEQ ID NO: 157),and RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZRNVPPIFNDVYWIAFCRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZRNVPPIFNDVYWIAFCRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZRNVPPIFNDVYWIAFZRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGIZRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZ ILMQYIKANSKFIGICRFRGLISLSQVYLS(SEQID NO: 158). In this case, the “a” denotes D-form alanine, the “Z”denotes 6-aminohexanoic acid, and the “(Cha)” denotesL-cyclohexylalanine.

Designing of Cyclic Form

The peptide may be designed to form a cyclic form. When the peptide isin a cyclic form, the stability in the body of a subject is increased;therefore, an improved effect can be expected when the peptide with acyclic form is used as an immunotherapeutic. In an embodiment, withregard to the peptide designed by a design method selected from thegroup consisting of a one unit design, a concatemer design, astring-of-beads design, and a mixed design method, the peptide may bedesigned to further have an auxiliary part having a function for forminga cyclic form at the N-terminus and C-terminus. In another embodiment,with regard to the peptide designed by a design method selected from thegroup consisting of a one unit design, a concatemer design, astring-of-beads design, and a mixed design method, the peptide may bedesigned to further include an auxiliary part and to form a cyclic formthrough the auxiliary part.

Other Designs

The peptide may be designed by other methods as necessary, in additionto the design methods described above. In an embodiment, with regard tothe peptide designed by a design method selected from the groupconsisting of a one unit design, a concatemer design, a string-of-beadsdesign, and a mixed design method, the peptide may further include oneor more auxiliary parts, one or more B-cell epitopes, and/or one or moreTh epitopes.

Sequences Similar to Peptide Units and/or Peptides Disclosed

In the present specification, peptide units and/or peptide which have asequence similar to those disclosed in the paragraphs of “unit-Adesign”, “unit-B design”, “unit-C design”, “unit-D design”, “unit-Edesign”, and “peptide design” above are disclosed.

In an embodiment, the peptide unit may have a sequence having anidentity of 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% to any one of thesequences disclosed in the paragraphs of “unit-A design”, “unit-Bdesign”, “unit-C design”, “unit-D design”, and “unit-E design”. Inanother embodiment, the peptide unit may have a sequence that matcheswith any one of the sequences disclosed in the paragraphs of “unit-Adesign”, “unit-B design”, “unit-C design”, “unit-D design”, and “unit-Edesign” by at least a numerical value selected in the immediatelypreceding sentence. In still another embodiment, the peptide unit mayhave a sequence which has an identity of 80%, 81%, 82%, 83%, 84%, 85%,86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or100% to any one selected from SEQ ID NO: 56 to SEQ ID NO: 150, SEQ IDNO: 160 to SEQ ID NO: 161, and SEQ ID NO: 198 to SEQ ID NO: 220. Instill another embodiment, the peptide unit may have a sequence whichmatches with any one of the sequences disclosed selected from SEQ ID NO:56 to SEQ ID NO: 150, SEQ ID NO: 160 to SEQ ID NO: 161, and SEQ ID NO:198 to SEQ ID NO: 220 by at least a numerical value selected in theimmediately preceding sentence. For example, the peptide unit may have asequence which has an identity of 90% or more to SEQ ID NO: 56.

In still another embodiment, the peptide may have a sequence which hasan identity of 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% to any one of thesequences disclosed in the paragraph of “peptide design”. In stillanother embodiment, the peptide may have a sequence that matches withany one of the sequences disclosed in the paragraph of “peptide design”by at least a numerical value selected in the immediately precedingsentence. In still another embodiment, the peptide may have a sequencewhich has an identity of 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%,89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% to anyone selected from SEQ ID NO: 151 to SEQ ID NO: 158. In still anotherembodiment, the peptide may have a sequence that matches with any one ofSEQ ID NO: 151 to SEQ ID NO: 158 by at least a numerical value selectedin the immediately preceding sentence. For example, the peptide may havea sequence which has an identity of 90% or more to SEQ ID NO: 151.

Use of Peptides

Use of Peptides—Overview

The peptides provided herein are suitable as an immunotherapeuticbecause they have the following characteristics when introduced into thebody of a subject: 1) they induce the production of antibodies thatspecifically bind to intentionally designed B-cell epitopes, and 2) theyinduce the production of uniform antibodies. Therefore, the peptides canbe used as an immunotherapeutic. In an embodiment, the peptides providedherein can be used as an immunotherapeutic for obesity. In anotherembodiment, the peptide units and/or peptides including the sameprovided herein may be used for the treatment of obesity.

Use of Peptides for Treatment of Obesity

The peptides provided herein include a B-cell epitope. In an embodiment,the B-cell epitope may be a B-cell epitope included in SEQ ID NOS: 41 to75. In particular, the

B-cell epitope is known to induce antibodies having an ability to bindto ApoB-100 (U.S. patent application Ser. No. 10/378,707,PCT/KR2005/000784, and Kim et al., 2016, An apolipoprotein B100 mimotopeprevents obesity in mice, Clinical Science 130, 105-116). It is knownfrom the prior documents above that when an antibody having an abilityto bind to ApoB-100 is induced by the B-cell epitope in the body of asubject, it has an immunotherapeutic effect on obesity. Accordingly, inthe present specification, the uses of the peptides for the treatment ofobesity and a method thereof are disclosed. In order to describe theimmunotherapeutic effects of the peptides on obesity, U.S. patentapplication Ser. No. 10/378,707, PCT/KR2005/000784, and Kim et al.,2016, An apolipoprotein B100 mimotope prevents obesity in mice, ClinicalScience 130, 105-116 are incorporated herein by reference. In the eventof a conflict between the referenced part and the description of thepresent specification, it should be construed that the description inthe present specification takes precedence over the referenced part.

Pharmaceutical Composition Including Peptide

The present specification discloses a pharmaceutical compositionincluding the peptide described above. The peptide can be used as animmunotherapeutic and has in common with vaccines in that it induces ahumoral immunity when injected into the body. Therefore, those skilledin the art may include, in the pharmaceutical composition including thepeptide, an appropriate constitution that can be added foradministration of general vaccines and/or to enhance the effect ofinducing an immune response. For example, the pharmaceutical compositionmay include the formulated peptide, pharmaceutically acceptablecarriers, supplements and/or adjuvants, but are not limited thereto.Specifically, the pharmaceutical composition may include water, saline,dextrose, ethanol, glycerol, sodium chloride, dextrose, mannitol,sorbitol, lactose, gelatin, albumin, aluminum hydroxide, Freund'sIncomplete Adjuvant and Complete Adjuvant (Pifco Laboratories, Detroit,Mich.), Merck adjuvant 65(Merck and Company, Inc., Rahway, N.J.),aluminum hydroxide gel (Alum), or aluminum salts such as aluminumphosphate, AS04 series, MF, squalene, MF59, QS21, calcium, iron or zincsalts, insoluble suspensions of acylated tyrosine, acylated fructose,cationically or anionically derived poly saccharides, polyphosphazenes,biodegradable microspheres, Quil A, toll-like receptor (TLR) agonists,PHAD [Avanti polar lipid, Monophosphoryl Lipid A (synthetic)],monophosphoryl lipid A (MPL, monophosphoryl Lipid A), synthetic lipid A,lipid A mimics or analogues, aluminum salts, cytokines, saponins,prolactin, growth hormone deoxycholic acid, betaglucan,polyribonucleotides, muramyl dipeptide (MDP) derivatives, CpG oligos,gram-negative bacterial lipopolysaccharide (LPS), polyphosphazene,emulsion, virosome, cochleate, poly(lactide-co-glycolide) (PLG)microparticles, poloxamer particles, microparticles, liposomes, or anappropriate combination thereof.

Manufacturing Method of the Peptides

The peptides provided herein may be prepared by a known method that canbe adopted by those skilled in the art, and the preparation method isnot particularly limited. In an embodiment, the peptide may be preparedby a recombinant protein preparation method. In another embodiment, thepeptide may be chemically synthesized. Specifically, the peptide may besynthesized by a liquid-phase peptide synthesis method, a solid-phasepeptide synthesis method, or a convergent method of small peptidefragments, but the methods are not limited thereto.

Nucleic Acid Encoding Peptide Unit and/or Peptide

Nucleic Acid Encoding Peptide Unit and/or Peptide—Overview

In the present specification, nucleic acids which encode peptide unitsand/or peptides disclosed above (hereinafter, “encoding nucleic acid”)are disclosed. While the peptide units and peptides disclosed herein mayinclude nonstandard amino acids, there are no codons corresponding tothe nonstandard amino acids in nature. Therefore, the nonstandard aminoacids cannot be encoded by a general method. Accordingly, it isnecessary to replace these nonstandard amino acids with appropriatestandard amino acids to encode them in the form of nucleic acids. Whenthe peptide unit and/or peptide do not include a nonstandard amino acid,the encoding nucleic acid can be designed using a nucleic acid codoncorresponding to each standard amino acid.

For convenience of description, in the present specification, thepeptide unit and/or peptide including the substitution of thenonstandard amino acid with an appropriate standard amino acid; and thepeptide unit and/or peptide including only standard amino acids arereferred to as a target peptide to be encoded, and the DNA and/or RNAencoding the target peptide to be encoded is referred to as an encodingnucleic acid. In particular, when the peptide unit and/or peptide do notinclude a nonstandard amino acid, the peptide unit and/or peptide hasthe same amino acid sequence as the target peptide to be encoded.

As used herein, the term “target peptide to be encoded” is a conceptualterm introduced to easily describe the resultant encoding nucleic acid,and is independent of the method or procedure for preparing the encodingnucleic acid.

Design of Target Peptide to be Encoded

When a peptide unit and peptide disclosed herein include a nonstandardamino acid, the target peptide to be encoded is designed by replacingthe same with an appropriate standard amino acid. When a peptide unitand peptide disclosed herein do not include a nonstandard amino acid,the corresponding target peptide to be encoded has the same sequence asthe peptide unit and peptide. In an embodiment, the target peptide to beencoded may be one in which the nonstandard amino acid is replaced withany standard amino acid. In another embodiment, the target peptide to beencoded may be one in which the nonstandard amino acid is replaced witha standard amino acid that is identical to the same or has an equivalentfunction. In still another embodiment, the target peptide to be encodedmay have a same sequence as the peptide unit and/or the peptide. Inparticular, the peptide unit and/or peptide are characterized by havingno nonstandard amino acids.

Embodiments of Designing Target Peptide to be Encoded—Cases IncludingPADRE

The peptide unit and peptide disclosed herein include one or more Thepitopes. In particular, when the Th epitope is a sequence named PADREdisclosed in U.S. patent application Ser. No. 305,871, it may include anonstandard amino acid, L-cyclohexylalanine. According to theliterature, the L-cyclohexylalanine may have both a function ofprotecting the PADRE from a peptide degrading enzyme and a function ofan anchor residue capable of binding to MHC Class II. Therefore, thetarget peptide to be encoded is designed by replacing theL-cyclohexylalanine with an appropriate standard amino acid having thesame or equivalent function. In an embodiment, the L-cyclohexylalaninemay be substituted with any standard amino acid. In another embodiment,the L-cyclohexylalanine may be substituted with phenylalanine ortyrosine. In still another embodiment, the sequence of the targetpeptide to be encoded corresponding to the PADRE may be KFVAAWTLKAA (SEQID NO: 195), KYVAAWTLKAA (SEQ ID NO: 196), or KXVAAWTLKAA (SEQ ID NO:197), in which the X refers to any standard amino acid.

Embodiments of Sequence of Target Peptide to be Encoded

In an embodiment, a sequence of the target peptide to be encoded isselected from RNVPPIFNDVYWIAFXXKXVAAWTLKAAXXCRFRGLISLSQVYLS(SEQ ID NO:198), RNVPPIFNDVYWIAFXXKXVAAWTLKAAXXGSHHHHHHGSDDDDK(SEQ ID NO: 199),GSHHHHHHGSDDDDKXXKXVAAWTLKAAXXRNVPPIFNDVYWIAF(SEQ ID NO: 200),KTTKQSFDLSVKAQYKKNKHXXKXVAAWTLKAAXXCRFRGLISLSQVYLS(SEQ ID NO: 201),RNVPPIFNDVYWIAFCRFRGLISLSQVYLSXXK(Cha)VAAWTLKAAXX(SEQ ID NO: 202),RNVPPIFNDVYWIAFXPKYVKQNTLKLATXCRFRGLISLSQVYLS(SEQ ID NO: 203),RNVPPIFNDVYWIAFXXKXVAAWTLKAAXX(SEQ ID NO: 204),RNVPPIFNDVYWIAFKXVAAWTLKAA(SEQ ID NO: 205),RNVPPIFNDVYWIAFKXVAAWTLKAAHHHHHH(SEQ ID NO: 206),RNVPPIFNDVYWIAFXXKXVAAWTLKAACRFRGLISLSQVYLS(SEQ ID NO: 207),RNVPPIFNDVYWIAFXXKXVAAWTLKAACR(SEQ ID NO: 208),RNVPPIFNDVYWIAFXXKFVAAWTLKAAXXCRFRGLISLSQVYLS(SEQ ID NO: 209),RNVPPIFNDVYWIAFXXKFVAAWTLKAAXX(SEQ ID NO: 210),RNVPPIFNDVYWIAFXXKFVAAWTLKAACRFRGLISLSQVYLS(SEQ ID NO: 211), andRNVPPIFNDVYWIAFXXKFVAAWTLKAACR(SEQ ID NO: 212). In this case, the “X”denotes any standard amino acid.

Design of Encoding Nucleic Acid 1—Nucleic Acid Codon Based

The encoding nucleic acid disclosed herein refers to a nucleic acidcodon encoding the encoding target peptide. Since the sequences of thetarget peptides to be encoded are all standard amino acids, the encodingnucleic acid is designed based on the nucleic acid codon correspondingto each amino acid of the target peptide to be encoded. In particular,since one or more nucleic acid codons can correspond to one standardamino acid, two or more encoding nucleic acids encoding one targetpeptide to be encoded can eventually be designed. In an embodiment, theencoding nucleic acid may be a DNA and/or RNA codon encoding the targetpeptide to be encoded. In another embodiment, the encoding nucleic acidmay have a DNA and/or RNA sequence, which is capable of a complementarybinding to a DNA and/or RNA codon encoding the target peptide to beencoded.

The sequence of the encoding nucleic acid may be codon optimized, whichis described in more detail below.

Design of Encoding Nucleic Acid 2—Codon Optimization

As described above, if the encoding nucleic acid is designed by simplylinking the nucleic acid codon corresponding to each amino acid of thetarget peptide to be encoded, multiple encoding nucleic acids may bedesigned for one target nucleic acid to be encoded. Since 1 to 6 nucleicacid codons correspond per standard amino acid on average, the number ofpossible nucleic acid codon combinations increases exponentially as theamino acid sequence length increases. However, not all of thesecombinations are of equal importance. In general, there is a combinationof nucleic acid codons capable of better expressing the target peptideto be encoded in cells, the combination may vary depending on thehigher-order structure of the sequence itself, the type of the targetcell into which the encoding nucleic acid is to be injected, etc.Discovering a combination of such a nucleic acid codon and specifyingthe discovered combination as a sequence of an encoding nucleic acid iscalled codon optimization. There is not necessarily only onecodon-optimized sequence for one coding target peptide, and there may betwo or more codon-optimized sequences.

In an embodiment, the encoding nucleic acid may have a codon optimizedDNA and/or RNA sequence. In another embodiment, the encoding nucleicacid may have non-codon optimized DNA and/or RNA sequences.

Codon Optimization of Encoding Nucleic Acid 1—Consideration ofHigher-Order Structure of Encoding Nucleic Acid

Codon optimization of the encoding nucleic acid may be performed inconsideration of a higher-order structure of the nucleic acid sequenceitself. In an embodiment, the encoding nucleic acid may becodon-optimized in consideration of the GC contents of the sequence. Inanother embodiment, the sequence of the encoding nucleic acid may have aGC content in the range of about less than 1%, about 1%, about 2%, about3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about10%, about 11%, about 12%, about 13%, about 14%, about 15%, about 16%,about 17%, about 18%, about 19%, about 20%, about 21%, about 22%, about23%, about 24%, about 25%, about 26%, about 27%, about 28%, about 29%,about 30%, about 31%, about 32%, about 33%, about 34%, about 35%, about36%, about 37%, about 38%, about 39%, about 40%, about 41%, about 42%,about 43%, about 44%, about 45%, about 46%, about 47%, about 48%, about49%, about 50%, about 51%, about 52%, about 53%, about 54%, about 55%,about 56%, about 57%, about 58%, about 59%, about 60%, about 61%, about62%, about 63%, about 64%, about 65%, about 66%, about 67%, about 68%,about 69%, about 70%, about 71%, about 72%, about 73%, about 74%, about75%, about 76%, about 77%, about 78%, about 79%, about 80%, about 81%,about 82%, about 83%, about 84%, about 85%, about 86%, about 87%, about88%, about 89%, about 90%, about 91%, about 92%, about 93%, about 94%,about 95%, about 96%, about 97%, about 98%, about 99%, or about 100%. Instill another embodiment, the sequence of the encoding nucleic acid mayhave a GC content within the numerical range selected in the immediatelypreceding sentence. For example, the sequence of the encoding nucleicacid may have a GC content in the range of about 20% to about 50%. Instill another embodiment, the sequence of the encoding nucleic acid mayhave a GC content less than the value selected in the immediatelypreceding sentence. For example, the sequence of the encoding nucleicacid may have a GC content of less than about 25%.

Codon Optimization of Encoding Nucleic Acid 2—Consideration of TargetCells to be Expressed

Codon optimization of the encoding nucleic acid may be achieved inconsideration of into which cell the encoding nucleic acid is to beinjected and expressed. In an embodiment, the codon optimization of theencoding nucleic acid may be achieved in consideration of the codonusage in prokaryotic or eukaryotic cells. In another embodiment, thecodon optimization of the encoding nucleic acid may be achieved inconsideration of codon usage of animal cells. In still anotherembodiment, the codon optimization of the encoding nucleic acid may beachieved in consideration of mammalian codon usage. In still anotherembodiment, the codon optimization of the encoding nucleic acid may beachieved in consideration of human codon usage. In still anotherembodiment, the encoding nucleic acid may be E. coli codon optimizedone. In still another embodiment, the encoding nucleic acid may bemammalian codon optimized one. In still another embodiment, the encodingnucleic acid may be human codon optimized one.

Embodiments of Encoding Nucleic Acid Sequences

In an embodiment, the encoding nucleic acid may be represented by asequence selected from5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGNNNGTGGCAGCTTGGAC CC TGAAGGCAGCANNNNNNTGCCGTTTCCGTGGACTGATTTCCCTGTCCCAGGTTTATCTGTCC-3′ (SEQ ID NO: 248),5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGTTCGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNNTGCCGTTTCCGTGGACTGATTTCCCTGTCCCAGGTTTATCTGTCC-3′ (SEQ ID NO: 249), 495′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGTATGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNNTGCCGTTTCCGTGGACTGATTTCCCTGTCCCAGGTTTATCTGTCC-3′ (SEQ ID NO: 250),5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGNNNGTGGCAGCTTGGACCCTGAAGGCANNNNNNGCAGGATCGCATCACCATCACCATCACGGATCCGATGATGATGACAAG-3′ (SEQ ID NO: 251),5′-ACGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGTTCGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNNGGATCGCATCACCATCACCATCACGGATCCGATGATGATGACAAG-3′ (SEQ ID NO: 252),5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGTATGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNNGGATCGCATCACCATCACCATCACGGATCCGATGATGATGACAAG-3′ (SEQ ID NO: 253),5′-GGATCGCATCACCATCACCATCACGGATCCGATGATGATGACAAGNNNNNNAAGNNNGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNNCGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTC-3′ (SEQ ID NO: 254),5′-GGATCGCATCACCATCACCATCACGGATCCGATGATGATGACAAGNNNNNNAAGTTCGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNNCGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTC-3′ (SEQ ID NO: 255),5′-GGATCGCATCACCATCACCATCACGGATCCGATGATGATGACAAGNNNNNNAAGTATGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNNCGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTC-3′ (SEQ ID NO: 256),5′-AAAACGACAAAGCAATCATTTGATTTAAGTGTAAAAGCTCAGTATNNNNNNAAGNNNGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNNTGCCGTTTCCGTGGACTGATTTCCCTGTCCCAGGTTTATCTGTCC-3′ (SEQ ID NO: 257),5′-AAAACGACAAAGCAATCATTTGATTTAAGTGTAAAAGCTCAGTATNNNNNNAAGTTCGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNNTGCCGTTTCCGTGGACTGATTTCCCTGTCCCAGGTTTATCTGTCC-3′ (SEQ ID NO: 258),5′-AAAACGACAAAGCAATCATTTGATTTAAGTGTAAAAGCTCAGTATNNNNNNAAGTATGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNNTGCCGTTTCCGTGGACTGATTTCCCTGTCCCAGGTTTATCTGTCC-3′ (SEQ ID NO: 259),5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCTGCCGTTTCCGTGGACTGATTTCCCTGTCCCAGGTTTATCTGTCCNNNNNNAAGNNNGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNN-3′ (SEQ ID NO: 260),5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCTGCCGTTTCCGTGGACTGATTTCCCTGTCCCAGGTTTATCTGTCCNNNNNNAAGTTCGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNN-3′ (SEQ ID NO: 261),5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCTGCCGTTTCCGTGGACTGATTTCCCTGTCCCAGGTTTATCTGTCCNNNNNNAAGTATG 50TGGCAGCTTGGACCCTGAAGGCAGCANNNNNN-3′ (SEQ ID NO: 262),5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNCCTAAGTATGTGAAGCAGAATACACTGAAGCTGGCAACCNNNTGCCGTTTCCGTGGACTGATTTCCCTGTCCCAGGTTTATCTGTCC-3′ (SEQ ID NO: 263),5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGNNNGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNN-3′ (SEQ ID NO: 264),5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGTTCGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNN-3′ (SEQ ID NO: 265),5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGTATGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNN-3′ (SEQ ID NO: 266),5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCAAGNNNGTGGCAGCTTGGACCCTGAAGGCAGCA-3′ (SEQ ID NO: 267),5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCAAGTTCGTGGCAGCTTGGACCCTGAAGGCAGCA-3′ (SEQ ID NO: 268),5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCAAGTATGTGGCAGCTTGGACCCTGAAGGCAGCA-3′ (SEQ ID NO: 269),5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCAAGNNNGTGGCAGCTTGGACCCTGAAGGCAGCACATCACCATCACCATCAC-3′ (SEQ ID NO: 270),5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCAAGTTCGTGGCAGCTTGGACCCTGAAGGCAGCACATCACCATCACCATCAC-3′ (SEQ ID NO: 271),5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCAAGTATGTGGCAGCTTGGACCCTGAAGGCAGCACATCACCATCACCATCAC-3′ (SEQ ID NO: 272),5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGNNNGTGGCAGCTTGGACCCTGAAGGCAGCATGCCGTTTCCGTGGACTGATTTCCCTGTCCCAGGTTTATCTGTCC-3′ (SEQ ID NO: 273),5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGTTCGTGGCAGCTTGGACCCTGAAGGCAGCATGCCGTTTCCGTGGACTGATTTCCCTGTCCCAGGTTTATCTGTCC-3′ (SEQ ID NO: 274),5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGTATGTGGCAGCTTGGACCCTGAAGGCAGCATGCCGTTTCCGTGGACTGATTTCCCTGTCCCAGGTTTATCTGTCC-3′ (SEQ ID NO: 275),5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNN 51NAAGNNNGTGGCAGCTTGGACCCTGAAGGCAGCATGCCGT-3′ (SEQ ID NO: 276),5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGTTCGTGGCAGCTTGGACCCTGAAGGCAGCATGCCGT-3′ (SEQ ID NO: 277), and5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGTATGTGGCAGCTTGGACCCTGAAGGCAGCATGCCGT-3′ (SEQ ID NO: 278).

In an embodiment, the encoding nucleic acid may be represented by an RNAsequence equivalent to the sequence selected from SEQ ID NO: 248 to SEQID NO: 278.

In another embodiment, the encoding nucleic acid may be one in which atleast one codon is substituted with a codon encoding the same amino acidin the sequence selected from SEQ ID NO: 248 to SEQ ID NO: 278. Forexample, the encoding nucleic acid may be one in which the first codon(the first to third nucleic acids at the 5′ end) in the SEQ ID NO: 248sequences (i.e., CGT) is substituted with CGC, CGG, CGA, AGA, or AGG.

In still another embodiment, the encoding nucleic acid may berepresented by an RNA sequence equivalent to that in the sequenceselected from SEQ ID NO: 248 to SEQ ID NO: 278, in which one or morecodons are substituted with a codon encoding the same amino acid.

Pharmaceutical Composition Including Nucleic Acid Encoding Peptide Unitand/or Peptide

Pharmaceutical Composition Including Encoding Nucleic Acid—Overview

The present specification provides a pharmaceutical compositionincluding a nucleic acid encoding a peptide unit and/or peptide (i.e.,encoding nucleic acid). In order to deliver the encoding nucleic acid toa subject to exhibit an intended effect of inducing an immune response,it needs to formulate the encoding nucleic acid by an appropriatemethod. The encoding nucleic acid may be formulated by a known method,for example, methods disclosed in W. K. KIM (2019, mRNA vaccine—new erain vaccinology, BRIC View 2019-R11), Zhang et al. (2019, Advances inmRNA Vaccines for Infectious Diseases, Frontiers in Immunology, Vol. 10,Article 594), Reichmuth et al. (2016, mRNA vaccine delivery using lipidnanoparticles, Therapeutic Delivery, 7(5), 319-334), Miao et al. (2021,mRNA vaccine for cancer immunotherapy, Molecular Cancer, 20:41), Boen etal. (2021, Identification of T Cell Ligands in a Library of PeptidesCovalently Attached to HLA-DR4, The Journal of Immunology,165:2040-2047), Pardi et al. (2018, mRNA vaccines—a new era invaccinology, Nature Reviews, Vol. 17, 261-279) and Korean PatentApplication No. 10-2017-0054429, but the method is not limited thereto.

The pharmaceutical composition including the encoding nucleic acid mayfurther include adjuvants and/or additional ingredients, in addition tothe formulated encoding nucleic acid. In an embodiment, thepharmaceutical composition including the encoding nucleic acid includesthe following: formulated encoding nucleic acids; optionally, adjuvants;and optionally, additional ingredients.

Formulated Encoding Nucleic Acid

The formulated encoding nucleic acid may be formulated by those skilledin the art by selecting an appropriate delivery means (vector) for theencoding nucleic acid. The encoding nucleic acid may be formulated usinga viral vector and/or a non-viral vector. In an embodiment, theformulated encoding nucleic acid may include a viral vector. In anotherembodiment, the formulated encoding nucleic acid may include a non-viralvector. Specifically, the non-viral vector may include lipids, polymers,and inorganic nanoparticles, but is not limited thereto.

In still another embodiment, the formulated encoding nucleic acid mayinclude one or more selected from the following:

a naked nucleic acid; a cationic peptide-complex nucleic acid(protamine); positively-charged oil-water cationic nanoemulsion(cationic nanoemulsion); a nucleic acid which is bound to chemically amodified dendrimer, and complexed with polyethylene glycol andPEG-lipids (modified dendrimer nanoparticle); a nucleic acid complexedwith protamine in PEG-lipid nanoparticles (protamine liposome); anucleic acid complexed with a cationic polymer (e.g., polyethylenimine(PEI)) (cationic polymer); a nucleic acid complexed with cationicpolymers such as PEI and lipid components(cationic polymer liposomes); anucleic acid complexed with a polysaccharide polymer (e.g., chitosan)(polysaccharide particles); a nucleic acid complexed with cationic lipidnanoparticle polymers (cationic lipid nanoparticle); a nucleic acidcomplexed with cationic lipid and cholesterol (cationiclipid-cholesterol nanoparticles); and a nucleic acid complexed withcationic lipid, cholesterol, and PEG-lipid (cationiclipid-cholesterol-PEG nanoparticles).

In still another embodiment, the formulated encoding nucleic acid mayinclude lipid nanoparticles (LNPs). In the specific embodiment above,the lipid nanoparticles may be ionizable cationic lipids, phospholipids,cholesterol, and/or lipid-anchored polyethylene glycol. Specifically,the ionizable cationic lipid may be one or more selected from thefollowing: DLin-DMA; DLin-KC2-DMA; DLin-MC3-DMA; C12-200; cKK-E12;DLin-MC3-DMA derivative L319 (Alnylam and AlCana Technologies); C12-200and cKK-E12 derivative (Anderson Group); COVID-19 vaccine lipid ALC-0315and SM-102; TT3 and biodegradable derivative FTT5 (Dong's group);vitamin-derived lipids ssPalmE and VcLNP; A9 (Acuitas); L5 (Moderna);A18 Lipid; ATX Lipid (LUNAR® composition; Arcturus); and LP01 (IntelliaTherapeutics). Specifically, the phospholipid may be one or moreselected from the following:1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE); and1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC).

In a specific embodiment, the formulated encoding nucleic acid mayinclude a polymer-based delivery system. In the specific embodiment, thepolymer-based delivery system may include one or more selected from thefollowing: polyethyleneimine (PEI); polyamidoamine (PAMAM);polypropyleneimine; and a polymer-based dendrimer.

In still another embodiment, the formulated encoding nucleic acid mayinclude a peptide-based delivery system. In the specific embodiment, thepeptide-based delivery system may include protamine. Specifically, theformulated encoding nucleic acid may be a protamine-mRNA complex.

In still another embodiment, the formulated encoding nucleic acid mayinclude cationic lipid constituting liposomes, lipoplexes and/orcationic emulsions (CNE). In the specific embodiment, the cationic lipidmay be 1,2-di-O-octadecenyl-3-trimethylammonium propane (DOTMA) and/or1,2-dioleoyl3-trimethylammonium-propane (DOTAP).

Adjuvants

In an embodiment, the pharmaceutical composition including the encodingnucleic acid may include lipid nanoparticles (LNPs), aluminum salts,1,2-dioleyl-3-trimethylammonium-propane chloride, MF59 (Novartis)adjuvant, CD70, CD40 ligand (CD40L), TriMix, protamine acting throughTLR7 signaling, and/or bacteria-derived monophosphoryl lipid A, asadjuvants.

Ingredients

In an embodiment, the pharmaceutical composition including the encodingnucleic acid may optionally include various additional ingredients. Inanother embodiment, the additional ingredients may be one or moreselected from the following:

lipids; salts to balance body acidity; sucrose to maintain stabilityduring repeated freezing-thawing; and vaccine stability enhancingsubstances

Specifically, the lipid may be SM-102, PEG2000-DMG, DPSC, cholesterol,and/or ALC-0315, but is not limited thereto. Specifically, the salt maybe sodium acetate, potassium chloride, monobasic potassium phosphate,sodium chloride, and/or dibasic sodium phosphate dehydrate, but is notlimited thereto. Specifically, the vaccine stability enhancing substancemay be acetic acid, an acid stabilizer (tromethamine), and/or ethanol,but is not limited thereto.

The Peptide—Summary

The peptide provided herein includes at least one peptide unit, and thepeptide unit includes at least one B-cell epitope and at least one Thepitope, and may include an appropriate number of auxiliary parts. Thepeptide unit is a part designed to uniformly induce only the intendedantibody while exhibiting a certain level of immunogenicity in the bodyof a subject. In addition, since the peptide unit is designed with arelatively short length, it has the characteristics of easy synthesisand a low production cost. The peptide has properties suitable for useas an immunotherapeutic due to the characteristics of the peptide unitdescribed above. In the present specification, the design principles ofthe peptide and the peptide unit are disclosed in detail.

The names for each part of the peptide disclosed herein (e.g., anauxiliary part) are given for convenience of explanation. Accordingly,the scope and name for each part may vary depending on the viewpoint.For example, the auxiliary part may be referred to as a protective part,a dummy part, and/or a linker, but is not limited thereto. For anotherexample, the B-cell epitope may be referred to as aTh-epitope-protective epitope, but is not limited thereto.

POSSIBLE EXAMPLES OF THE INVENTION

Hereinafter, possible examples of the invention provided in the presentspecification are listed. The following Examples provided in thisparagraph merely correspond to embodiments of the invention. Therefore,the invention provided in the present specification cannot beinterpreted as being limited to the following examples.

Symbols Used in Each Example

Hereinafter, symbols used for a brief description of each Example, inaddition to numbers to distinguish each Example, will be described.

“B” denotes a B-cell epitope. “T” denotes Th epitope. “A” denotes anauxiliary part (Auxiliary part). “U” denotes a peptide unit.

When each component is linked by “-”, the component means that thecomponents on either side of the “-” are directly linked or linkedthrough any other component. For example, when it is described as B-T,it includes all of the peptides in which a B-cell epitope and a Thepitope are directly linked, and a peptide in which a B-cell epitope anda Th epitope are linked via any other sequence.

If necessary, each component may be marked with a subscript number,representing that the two components are different. For example, whenexpressed as B1-B2-T, B1 and B2 represent different B-cell epitopes.

The above symbols are only those for schematically describing Examples,and they should not be interpreted by limiting Examples with thesesymbols.

Peptide Unit 1

Example 1 Peptide Unit that Can Induce Humoral Immunity

A peptide unit that can induce a humoral immunity by being recognized byCD4+ T-cells, including the following: at least one Th epitope; and atleast one B-cell epitope, in which the peptide unit is characterized inthat the length of the Th epitope is 8mer, 9mer, 10mer, 11mer, 12mer,13mer, 14mer, 15mer, 16mer, 17mer, 18mer, 19mer, 20mer, 21mer, 22mer,23mer, 24mer, 25mer, 26mer, 27mer, 28mer, 29mer, or 30mer; the length ofthe B-cell epitope is 8mer, 9mer, 10mer, 11mer, 12mer, 13mer, 14mer,15mer, 16mer, 17mer, 18mer, 19mer, 20mer, 21mer, 22mer, 23mer, 24mer,25mer, 26mer, 27mer, 28mer, 29mer, 30mer, 31mer, or 32mer; and thelength of the peptide unit is 16mer, 17mer, 18mer, 19mer, 20mer, 21mer,22mer, 23mer, 24mer, 25mer, 26mer, 27mer, 28mer, 29mer, 30mer, 31mer,32mer, 33mer, 34mer, 35mer, 36mer, 37mer, 38mer, 39mer, 40mer, 41mer,42mer, 43mer, 44mer, 45mer, 46mer, 47mer, 48mer, 49mer, 50mer, 51mer,52mer, 53mer, 54mer, 55mer, 56mer, 57mer, 58mer, 59mer, 60mer, 61mer,62mer, 63mer, 64mer, 65mer, 66mer, 67mer, 68mer, 69mer, 70mer, 71mer,72mer, 73mer, 74mer, 75mer, 76mer, 77mer, 78mer, 79mer, 80mer, 81mer,82mer, 83mer, 84mer, 85mer, 86mer, 87mer, 88mer, 89mer, 90mer, 91mer,92mer, 93mer, 94mer, 95mer, 96mer, 97mer, 98mer, 99mer, or 100mer.

Example 2 Peptide Unit that Induces Antibodies Targeting ApolipoproteinB-100

A peptide unit that can induce a humoral immunity by being recognized byCD4+ T-cells, including the following: at least one Th epitope; and atleast one B-cell epitope, in which the peptide unit is characterized inthat the length of the Th epitope is 8mer, 9mer, 10mer, 11mer, 12mer,13mer, 14mer, 15mer, 16mer, 17mer, 18mer, 19mer, 20mer, 21mer, 22mer,23mer, 24mer, 25mer, 26mer, 27mer, 28mer, 29mer, or 30mer; the B-cellepitope is able to induce antibodies that target apolipoprotein B-100;and the length of the peptide unit is 16mer, 17mer, 18mer, 19mer, 20mer,21mer, 22mer, 23mer, 24mer, 25mer, 26mer, 27mer, 28mer, 29mer, 30mer,31mer, 32mer, 33mer, 34mer, 35mer, 36mer, 37mer, 38mer, 39mer, 40mer,41mer, 42mer, 43mer, 44mer, 45mer, 46mer, 47mer, 48mer, 49mer, 50mer,51mer, 52mer, 53mer, 54mer, 55mer, 56mer, 57mer, 58mer, 59mer, 60mer,61mer, 62mer, 63mer, 64mer, 65mer, 66mer, 67mer, 68mer, 69mer, 70mer,71mer, 72mer, 73mer, 74mer, 75mer, 76mer, 77mer, 78mer, 79mer, 80mer,81mer, 82mer, 83mer, 84mer, 85mer, 86mer, 87mer, 88mer, 89mer, 90mer,91mer, 92mer, 93mer, 94mer, 95mer, 96mer, 97mer, 98mer, 99mer, or100mer.

Example 3 Peptide Unit that Induces Antibody which TargetsApolipoprotein B-100 Included in LDL and/or VLDL

The peptide unit of Example 2, wherein the B-cell epitope ischaracterized in that it induces an antibody targeting a site selectedfrom the following: an externally exposed site of apolipoprotein B-100included in low-density lipoprotein (LDL); and an externally exposedsite of apolipoprotein B-100 included in ultra-low-density lipoprotein(VLDL).

Example 4 Limitation on Type B

The peptide unit of Example 2, wherein the peptide unit is characterizedin that the B-cell epitope is a fragment of apolipoprotein B-100 and/ora mimotope of apolipoprotein B-100.

Example 5 Limitation on Sequence of B

The peptide unit of Example 4, wherein the peptide unit is characterizedin that the B-cell epitope is a sequence being selected from the groupconsisting of RNVPPIFNDVYWIAF (SEQ ID NO: 6), CRFRGLISLSQVYLS (SEQ IDNO: 7), KTTKQSFDLSVKAQYKKNKH (SEQ ID NO: 8), RFRGLISLSQVYLDP (SEQ ID NO:221), and SVCGCPVGHHDVVGL (SEQ ID NO: 222) or an epitope included in thesequence selected from SEQ ID NOS: 6 to 8 and 221 to 222.

Example 6 Limitation on Full-Length Sequence Length of Peptide Unit

The peptide unit of any one of Examples 1 to 5, wherein the peptide unitis characterized in that the length of the peptide unit is 23mer to71mer or 26mer to 50mer.

Peptide Unit 2—Unit A

Example 7 B-T and T-B

The peptide unit of any one of Examples 1 to 4, wherein the peptide unitis characterized in that the peptide unit includes one B-cell epitopeand one Th epitope.

Example 8 Limitation on Sequence B, Limitation on Length T, andLimitation on Length U

The peptide unit of Example 7, wherein the peptide unit is characterizedin that the length of the peptide unit is 26mer to 45mer; the B-cellepitope is selected from RNVPPIFNDVYWIAF (SEQ ID NO: 6), CRFRGLISLSQVYLS(SEQ ID NO: 7), KTTKQSFDLSVKAQYKKNKH (SEQ ID NO: 8), RFRGLISLSQVYLDP(SEQ ID NO: 221), and SVCGCPVGHHDVVGL (SEQ ID NO: 222); and the lengthof the Th epitope is 11mer to 13mer.

Example 9 Limitation on Sequence T

The peptide unit of any one of Examples 7 and 8, wherein the Th epitopeis characterized in that it is selected from the group consisting of thefollowing: K(Cha)VAAWTLKAA (SEQ ID NO: 1); PKYVKQNTLKLAT (SEQ ID NO: 2);ILMQYIKANSKFIGI (SEQ ID NO: 3); QSIALSSLMVAQAIP (SEQ ID NO: 4);ILMQYIKANSKFIGIPMGLPQSIALSSLMVAQ (SEQ ID NO: 5); PLGFFPDHQL (SEQ ID NO:162); WPEANQVGAGAFGPGF (SEQ ID NO: 163); MQWNSTALHQALQDP (SEQ ID NO:164); MQWNSTTFHQTLQDPRVRGLYFPAGG (SEQ ID NO: 165); FFLLTRILTI (SEQ IDNO: 166); FFLLTRILTIPQSLD (SEQ ID NO: 167); TSLNFLGGTTVCLGQ (SEQ ID NO:168); QSPTSNHSPTSCPPIC (SEQ ID NO: 169); IIFLFILLLCLIFLLVLLD (SEQ ID NO:170); CTTPAQGNSMFPSC (SEQ ID NO: 171); CTKPTDGN (SEQ ID NO: 172);WASVRFSW (SEQ ID NO: 173); LLPIFFCLW (SEQ ID NO: 174);MDIDPYKEFGATVELLSFLP (SEQ ID NO: 175); FLPSDFFPSV (SEQ ID NO: 176);RDLLDTASALYREALESPEH (SEQ ID NO: 177); PHHTALRQAILCWGELMTLA (SEQ ID NO:178); GRETVIEYLVSFGVW (SEQ ID NO: 179); EYLVSFGVWIRTPPA (SEQ ID NO:180); VSFGVWIRTPPAYRPPNAPI (SEQ ID NO: 181); TVVRRRGRSP (SEQ ID NO:182); VGPLTVNEKRRLKLI (SEQ ID NO: 183); RHYLHTLWKAGILYK (SEQ ID NO:184); ESRLVVDFSQFSRGN (SEQ ID NO: 185); LQSLTNLLSSNLSWL (SEQ ID NO:186); SSNLSWLSLDVSAAF (SEQ ID NO: 187); LHLYSHPIILGFRKI (SEQ ID NO:188); KQCFRKLPVNRPIDW (SEQ ID NO: 189); LCQVFADATPTGWGL (SEQ ID NO:190); AANWILRGTSFVYVP (SEQ ID NO: 191); and EIRLKVFVLGGCRHK (SEQ ID NO:192), in which the (Cha) denotes L-cyclohexylalanins.

Example 10 BAT and TAB

The peptide unit of any one of Examples 7 and 8, wherein the peptideunit is characterized in that it further includes an auxiliary part, andthat the auxiliary part is linked between the B-cell epitope and the Thepitope.

Example 11 Limitation on A, Including Nonstandard Amino Acid

The peptide unit of Example 10, wherein the peptide unit ischaracterized in that the auxiliary part includes one or morenonstandard amino acids.

Example 12 Limitation on Sequence of A Including Nonstandard Amino Acid

The peptide unit of Example 11, wherein the peptide unit ischaracterized in that the auxiliary part is selected from a, Z, aZ, Za,GSHHHHHHGSDDDKZa (SEQ ID NO: 193), and aZGSHHHHHHGSDDDK (SEQ ID NO:194).

Example 13 Limitation on Sequence A Not Including Nonstandard Amino Acid

The peptide unit of Example 10, wherein the peptide unit ischaracterized in that the auxiliary part is selected from CR, HHHHHH(SEQ ID NO: 53), and RRRRRR (SEQ ID NO: 159).

Peptide Unit 3—Unit D

Example 14 B-T₁-T₂ and T₂-T₁-B

The peptide unit of any one of Examples 1 to 4, wherein the peptide unitis characterized in that it includes one B-cell epitope, and two Thepitopes which are referred to as the first Th epitope and the second Thepitope, respectively, and that the first Th epitope is linked betweenthe B-cell epitope and the second Th epitope.

Example 15 Limitation on Sequence B, Limitation on Length T, andLimitation on Length U

The peptide unit of Example 14, wherein the peptide unit ischaracterized in that the length of the peptide unit is 37mer to 50mer;the B-cell epitope is selected from RNVPPIFNDVYWIAF (SEQ ID NO: 6),CRFRGLISLSQVYLS (SEQ ID NO: 7), KTTKQSFDLSVKAQYKKNKH (SEQ ID NO: 8),RFRGLISLSQVYLDP (SEQ ID NO: 221), and SVCGCPVGHHDVVGL (SEQ ID NO: 222);and the length of the first Th epitope and the second Th epitope is11mer to 13mer, respectively.

Example 16 Limitation on Sequence T

The peptide unit of any one of Examples 14 and 15, wherein the Thepitope is characterized in that it is selected from the groupconsisting of the following: K(Cha)VAAWTLKAA (SEQ ID NO: 1);PKYVKQNTLKLAT (SEQ ID NO: 2); ILMQYIKANSKFIGI (SEQ ID NO: 3);QSIALSSLMVAQAIP (SEQ ID NO: 4); ILMQYIKANSKFIGIPMGLPQSIALSSLMVAQ (SEQ IDNO: 5); PLGFFPDHQL (SEQ ID NO: 162); WPEANQVGAGAFGPGF (SEQ ID NO: 163);MQWNSTALHQALQDP (SEQ ID NO: 164); MQWNSTTFHQTLQDPRVRGLYFPAGG (SEQ ID NO:165); FFLLTRILTI (SEQ ID NO: 166); FFLLTRILTIPQSLD (SEQ ID NO: 167);TSLNFLGGTTVCLGQ (SEQ ID NO: 168); QSPTSNHSPTSCPPIC (SEQ ID NO: 169);IIFLFILLLCLIFLLVLLD (SEQ ID NO: 170); CTTPAQGNSMFPSC (SEQ ID NO: 171);CTKPTDGN (SEQ ID NO: 172); WASVRFSW (SEQ ID NO: 173); LLPIFFCLW (SEQ IDNO: 174); MDIDPYKEFGATVELLSFLP (SEQ ID NO: 175); FLPSDFFPSV (SEQ ID NO:176); RDLLDTASALYREALESPEH (SEQ ID NO: 177); PHHTALRQAILCWGELMTLA (SEQID NO: 178); GRETVIEYLVSFGVW (SEQ ID NO: 179); EYLVSFGVWIRTPPA (SEQ IDNO: 180); VSFGVWIRTPPAYRPPNAPI (SEQ ID NO: 181); TVVRRRGRSP (SEQ ID NO:182); VGPLTVNEKRRLKLI (SEQ ID NO: 183); RHYLHTLWKAGILYK (SEQ ID NO:184); ESRLVVDFSQFSRGN (SEQ ID NO: 185); LQSLTNLLSSNLSWL (SEQ ID NO:186); SSNLSWLSLDVSAAF (SEQ ID NO: 187); LHLYSHPIILGFRKI (SEQ ID NO:188); KQCFRKLPVNRPIDW (SEQ ID NO: 189); LCQVFADATPTGWGL (SEQ ID NO:190); AANWILRGTSFVYVP (SEQ ID NO: 191); and EIRLKVFVLGGCRHK (SEQ ID NO:192), in which the a denotes D-form alanine, the (Cha) denotesL-cyclohexylalanins, and the Z denotes 6-aminohexanoic acid.

Example 17 BAT₁-T₂ and T₂-T₁AB

The peptide unit of any one of Examples 14 and 15, wherein the peptideunit is characterized in that it further includes an auxiliary part, andthe auxiliary part is linked between the B-cell epitope and the first Thepitope.

Example 18 B-T₁AT₂ and T₂AT₁-B

The peptide unit of any one of Examples 14 and 15, wherein the peptideunit is characterized in that it further includes an auxiliary part, andthe auxiliary part is linked between the first Th epitope and the secondTh epitope.

Example 19 Limitation on A, Including Nonstandard Amino Acid

The peptide unit of any one of Examples 17 and 18, wherein the peptideunit is characterized in that the auxiliary part includes one or morenonstandard amino acids.

Example 20 Limitation on Sequence A Including Nonstandard Amino Acid

The peptide unit of Example 19, wherein the peptide unit ischaracterized in that the auxiliary part is selected from a, Z, aZ, Za,GSHHHHHHGSDDDKZa (SEQ ID NO: 193), and aZGSHHHHHHGSDDDK (SEQ ID NO:194).

Example 21 Limitation on Sequence A Not Including Nonstandard Amino Acid

The peptide unit of any one of Examples 17 and 18, wherein the peptideunit is characterized in that the auxiliary part is selected from CR,HHHHHH (SEQ ID NO: 53), and RRRRRR (SEQ ID NO: 159).

Example 22 BAT₁AT₂ and T₂AT₁AB

The peptide unit of any one of Examples 14 and 15, wherein the peptideunit is characterized in that it further includes a first auxiliary partand a second auxiliary part, and that the first auxiliary part is linkedbetween the B-cell epitope and the Th epitope and the second auxiliarypart is linked between the first Th epitope and the second Th epitope.

Example 23 Limitation on A, Including Nonstandard Amino Acid

The peptide unit of Example 22, wherein the peptide unit ischaracterized in that the first auxiliary part and/or the secondauxiliary part include one or more nonstandard amino acids.

Example 24 Limitation on Sequence of A Including Nonstandard Amino Acid

The peptide unit of Example 23, wherein the peptide unit ischaracterized in that the auxiliary part including one or morenonstandard amino acids is independently selected from a, Z, aZ, Za,GSHHHHHHGSDDDKZa (SEQ ID NO: 193), and aZGSHHHHHHGSDDDK (SEQ ID NO:194).

Example 25 Limitation on Sequence A Not Including Nonstandard Amino Acid

The peptide unit of Example 22, wherein the peptide unit ischaracterized in that the first auxiliary part and/or the secondauxiliary part are independently selected from CR, HHHHHH (SEQ ID NO:53), and RRRRRR (SEQ ID NO: 159).

Peptide Unit 4—Unit B

Example 26 B₁-B₂-T and T-B₂-B₁

The peptide unit of any one of Examples 1 to 4, wherein the peptide unitis characterized in that it includes two B-cell epitopes, which arereferred to as a first B-cell epitope and a second B-cell epitope,respectively, and that the second B-cell epitope is linked between thefirst B-cell epitope and the Th epitope.

Example 27 Limitation on Sequence B, Limitation on Length T, andLimitation on Length U

The peptide unit of Example 26, wherein the peptide unit ischaracterized in that the length of the peptide unit is 45mer to 50mer;the first B-cell epitope and the second B-cell epitope are independentlyselected from RNVPPIFNDVYWIAF (SEQ ID NO: 6), CRFRGLISLSQVYLS (SEQ IDNO: 7), KTTKQSFDLSVKAQYKKNKH (SEQ ID NO: 8), RFRGLISLSQVYLDP (SEQ ID NO:221), and SVCGCPVGHHDVVGL (SEQ ID NO: 222); and the length of the Thepitope is 11mer to 13mer.

Example 28 Limitation on Sequence T

The peptide unit of any one of Examples 26 and 27, wherein the Thepitope is characterized in that it is selected from the groupconsisting of the following: K(Cha)VAAWTLKAA (SEQ ID NO: 1);PKYVKQNTLKLAT (SEQ ID NO: 2); ILMQYIKANSKFIGI (SEQ ID NO: 3);QSIALSSLMVAQAIP (SEQ ID NO: 4); ILMQYIKANSKFIGIPMGLPQSIALSSLMVAQ (SEQ IDNO: 5); PLGFFPDHQL (SEQ ID NO: 162); WPEANQVGAGAFGPGF (SEQ ID NO: 163);MQWNSTALHQALQDP (SEQ ID NO: 164); MQWNSTTFHQTLQDPRVRGLYFPAGG (SEQ ID NO:165); FFLLTRILTI (SEQ ID NO: 166); FFLLTRILTIPQSLD (SEQ ID NO: 167);TSLNFLGGTTVCLGQ (SEQ ID NO: 168); QSPTSNHSPTSCPPIC (SEQ ID NO: 169);IIFLFILLLCLIFLLVLLD (SEQ ID NO: 170); CTTPAQGNSMFPSC (SEQ ID NO: 171);CTKPTDGN (SEQ ID NO: 172); WASVRFSW (SEQ ID NO: 173); LLPIFFCLW (SEQ IDNO: 174); MDIDPYKEFGATVELLSFLP (SEQ ID NO: 175); FLPSDFFPSV (SEQ ID NO:176); RDLLDTASALYREALESPEH (SEQ ID NO: 177); PHHTALRQAILCWGELMTLA (SEQID NO: 178); GRETVIEYLVSFGVW (SEQ ID NO: 179); EYLVSFGVWIRTPPA (SEQ IDNO: 180); VSFGVWIRTPPAYRPPNAPI (SEQ ID NO: 181); TVVRRRGRSP (SEQ ID NO:182); VGPLTVNEKRRLKLI (SEQ ID NO: 183); RHYLHTLWKAGILYK (SEQ ID NO:184); ESRLVVDFSQFSRGN (SEQ ID NO: 185); LQSLTNLLSSNLSWL (SEQ ID NO:186); SSNLSWLSLDVSAAF (SEQ ID NO: 187); LHLYSHPIILGFRKI (SEQ ID NO:188); KQCFRKLPVNRPIDW (SEQ ID NO: 189); LCQVFADATPTGWGL (SEQ ID NO:190); AANWILRGTSFVYVP (SEQ ID NO: 191); and EIRLKVFVLGGCRHK (SEQ ID NO:192), in which the a denotes D-form alanine, the (Cha) denotesL-cyclohexylalanins, and the Z denotes 6-aminohexanoic acid.

Example 29 B₁-B₂AT and TAB₂-B₁

The peptide unit of any one of Examples 26 and 27, wherein the peptideunit is characterized in that it further includes an auxiliary part, andthe auxiliary part is linked between the second B-cell epitope and theTh epitope.

Example 30 B₁AB₂-T and T-B₂AB₁

The peptide unit of any one of Examples 26 and 27, wherein the peptideunit is characterized in that it further includes an auxiliary part, andthe auxiliary part is linked between the first B-cell epitope and thesecond B-cell epitope.

Example 31 Limitation on A, Including Nonstandard Amino Acid

The peptide unit of any one of Examples 29 and 30, wherein the peptideunit is characterized in that the auxiliary part includes one or morenonstandard amino acids.

Example 32 Limitation on Sequence A Including Nonstandard Amino Acid

The peptide unit of Example 31, wherein the peptide unit ischaracterized in that the auxiliary part is selected from a, Z, aZ, Za,GSHHHHHHGSDDDKZa (SEQ ID NO: 193), and aZGSHHHHHHGSDDDK (SEQ ID NO:194).

Example 33 A, Limitation on Sequence Not Including Nonstandard AminoAcid

The peptide unit of any one of Examples 29 and 30, wherein the peptideunit is characterized in that the auxiliary part is selected from CR,HHHHHH (SEQ ID NO: 53), and RRRRRR (SEQ ID NO: 159).

Example 34 B₁A₁B₂A₂T and TA₂B₂A₁B₁

The peptide unit of any one of Examples 26 and 27, wherein the peptideunit is characterized in that it further includes a first auxiliary partand a second auxiliary part, and the first auxiliary part is linkedbetween the first B-cell epitope and the second B-cell epitope, and thesecond auxiliary part is linked between the second B-cell epitope and Thepitope.

Example 35 Limitation on A, Including Nonstandard Amino Acid

The peptide unit of Example 34, wherein the peptide unit ischaracterized in that the first auxiliary part and/or the secondauxiliary part include one or more nonstandard amino acids.

Example 36 Limitation on Sequence A Including Nonstandard Amino Acid

The peptide unit of Example 35, wherein the peptide unit ischaracterized in that the auxiliary parts including one or morenonstandard amino acids are each independently selected from a, Z, aZ,Za, GSHHHHHHGSDDDKZa (SEQ ID NO: 193), and aZGSHHHHHHGSDDDK (SEQ ID NO:194).

Example 37 Limitation on Sequence A Not Including Nonstandard Amino Acid

The peptide unit of Example 34, wherein the peptide unit ischaracterized in that the first auxiliary part and/or the secondauxiliary part are each independently selected from CR, HHHHHH (SEQ IDNO: 53), and RRRRRR (SEQ ID NO: 159).

Peptide Unit 5—Unit C

Example 38 B₁-T-B₂

The peptide unit of any one of Examples 1 to 4, wherein the peptide unitis characterized in that the peptide unit includes two B-cell epitopes,which are referred to as a first B-cell epitope and a second B-cellepitope, and one Th epitope, and the Th epitope is linked between thefirst B-cell epitope and the second B-cell epitope.

Example 39 Limitation on Sequence B, Limitation on Length T, andLimitation on Length U

The peptide unit of Example 38, wherein the peptide unit ischaracterized in that the length of the peptide unit is 45mer to 50mer;the first B-cell epitope and the second B-cell epitope are eachindependently selected from RNVPPIFNDVYWIAF (SEQ ID NO: 6),CRFRGLISLSQVYLS (SEQ ID NO: 7), KTTKQSFDLSVKAQYKKNKH (SEQ ID NO: 8),RFRGLISLSQVYLDP (SEQ ID NO: 221), and SVCGCPVGHHDVVGL (SEQ ID NO: 222);and the length of the Th epitope is 11mer to 13mer.

Example 40 Limitation on Sequence T

The peptide unit of any one of Examples 38 and 39, wherein the Thepitope is characterized in that it is selected from the groupconsisting of the following: K(Cha)VAAWTLKAA (SEQ ID NO: 1);PKYVKQNTLKLAT (SEQ ID NO: 2); ILMQYIKANSKFIGI (SEQ ID NO: 3);QSIALSSLMVAQAIP (SEQ ID NO: 4); ILMQYIKANSKFIGIPMGLPQSIALSSLMVAQ (SEQ IDNO: 5); PLGFFPDHQL (SEQ ID NO: 162); WPEANQVGAGAFGPGF (SEQ ID NO: 163);MQWNSTALHQALQDP (SEQ ID NO: 164); MQWNSTTFHQTLQDPRVRGLYFPAGG (SEQ ID NO:165); FFLLTRILTI (SEQ ID NO: 166); FFLLTRILTIPQSLD (SEQ ID NO: 167);TSLNFLGGTTVCLGQ (SEQ ID NO: 168); QSPTSNHSPTSCPPIC (SEQ ID NO: 169);IIFLFILLLCLIFLLVLLD (SEQ ID NO: 170); CTTPAQGNSMFPSC (SEQ ID NO: 171);CTKPTDGN (SEQ ID NO: 172); WASVRFSW (SEQ ID NO: 173); LLPIFFCLW (SEQ IDNO: 174); MDIDPYKEFGATVELLSFLP (SEQ ID NO: 175); FLPSDFFPSV (SEQ ID NO:176); RDLLDTASALYREALESPEH (SEQ ID NO: 177); PHHTALRQAILCWGELMTLA (SEQID NO: 178); GRETVIEYLVSFGVW (SEQ ID NO: 179); EYLVSFGVWIRTPPA (SEQ IDNO: 180); VSFGVWIRTPPAYRPPNAPI (SEQ ID NO: 181); TVVRRRGRSP (SEQ ID NO:182); VGPLTVNEKRRLKLI (SEQ ID NO: 183); RHYLHTLWKAGILYK (SEQ ID NO:184); ESRLVVDFSQFSRGN (SEQ ID NO: 185); LQSLTNLLSSNLSWL (SEQ ID NO:186); SSNLSWLSLDVSAAF (SEQ ID NO: 187); LHLYSHPIILGFRKI (SEQ ID NO:188); KQCFRKLPVNRPIDW (SEQ ID NO: 189); LCQVFADATPTGWGL (SEQ ID NO:190); AANWILRGTSFVYVP (SEQ ID NO: 191); and EIRLKVFVLGGCRHK (SEQ ID NO:192), in which the a denotes D-form alanine, the (Cha) denotesL-cyclohexylalanins, and the Z denotes 6-aminohexanoic acid.

Example 41 B₁AT-B₂ and B₂-TAB₁

The peptide unit of any one of Examples 38 and 39, wherein the peptideunit is characterized in that it further includes an auxiliary part, andthe auxiliary part is linked between the first B-cell epitope and the Thepitope.

Example 42 Limitation on A, Including Nonstandard Amino Acid

The peptide unit of Example 41, wherein the peptide unit ischaracterized in that the auxiliary part includes one or morenonstandard amino acids,

Example 43 Limitation on Sequence A Including Nonstandard Amino Acid

The peptide unit of Example 42, wherein the peptide unit ischaracterized in that the auxiliary part is selected from a, Z, aZ, Za,GSHHHHHHGSDDDKZa (SEQ ID NO: 193), and aZGSHHHHHHGSDDDK (SEQ ID NO:194).

Example 44 Limitation on Sequence A Not Including Nonstandard Amino Acid

The peptide unit of Example 41, wherein the peptide unit ischaracterized in that the auxiliary part is selected from CR, HHHHHH(SEQ ID NO: 53), and RRRRRR (SEQ ID NO: 159).

Example 45 B₁A₁TA₂B₂

The peptide unit of any one of Examples 38 and 39, wherein the peptideunit is characterized in that it further includes a first auxiliary partand a second auxiliary part; the first auxiliary part is linked betweenthe first B-cell epitope and the Th epitope; and the second auxiliarypart is linked between the second B-cell epitope and the Th epitope.

Example 46 Limitation on A, Including Nonstandard Amino Acid

The peptide unit of Example 45, wherein the peptide unit ischaracterized in that the first auxiliary part and/or the secondauxiliary part include one or more nonstandard amino acids.

Example 47 Limitation on Sequence A Including Nonstandard Amino Acid

The peptide unit of Example 46, wherein the peptide unit ischaracterized in that the auxiliary parts including one or morenonstandard amino acids are each independently selected from a, Z, aZ,Za, GSHHHHHHGSDDDKZa (SEQ ID NO: 193), and aZGSHHHHHHGSDDDK (SEQ ID NO:194).

Example 48 Limitation on Sequence A Not Including Nonstandard Amino Acid

The peptide unit of Example 45, wherein the peptide unit ischaracterized in that the first auxiliary part and/or the secondauxiliary part are each independently selected from CR, HHHHHH (SEQ IDNO: 53), and RRRRRR (SEQ ID NO: 159).

Peptide Unit 6—Unit E

Example 49 B₁-T₁-T₂-B₂

The peptide unit of any one of Examples 1 to 4, wherein the peptide unitis characterized in that it includes two B-cell epitopes, which are eachreferred to as a first B-cell epitope and a second B-cell epitope, andtwo Th epitopes, which are each referred to as a first Th epitope and asecond Th epitope, a first B-cell epitope, a first Th epitope, a secondTh epitope, and a second B-cell epitope are sequentially linked, in thedirection from the N-terminus to the C-terminus.

Example 50 Limitation on Sequence B, Limitation on Length T, andLimitation on Length U

The peptide unit of Example 49, wherein the peptide unit ischaracterized in that the length of the peptide unit is 52mer to 90mer;the first B-cell epitope and the second B-cell epitope are eachindependently selected from RNVPPIFNDVYWIAF (SEQ ID NO: 6),CRFRGLISLSQVYLS (SEQ ID NO: 7), KTTKQSFDLSVKAQYKKNKH (SEQ ID NO: 8),RFRGLISLSQVYLDP (SEQ ID NO: 221), and SVCGCPVGHHDVVGL (SEQ ID NO: 222);and the length of the first Th epitope and the second Th epitope areeach 11mer to 13mer.

Example 51 Limitation on Sequence T

The peptide unit of Example 50, wherein the first Th epitope and thesecond Th epitope are characterized in that they are each independentlyselected from the group consisting of the following: K(Cha)VAAWTLKAA(SEQ ID NO: 1); PKYVKQNTLKLAT (SEQ ID NO: 2); ILMQYIKANSKFIGI (SEQ IDNO: 3); QSIALSSLMVAQAIP (SEQ ID NO: 4); ILMQYIKANSKFIGIPMGLPQSIALSSLMVAQ(SEQ ID NO: 5); PLGFFPDHQL (SEQ ID NO: 162); WPEANQVGAGAFGPGF (SEQ IDNO: 163); MQWNSTALHQALQDP (SEQ ID NO: 164); MQWNSTTFHQTLQDPRVRGLYFPAGG(SEQ ID NO: 165); FFLLTRILTI (SEQ ID NO: 166); FFLLTRILTIPQSLD (SEQ IDNO: 167); TSLNFLGGTTVCLGQ (SEQ ID NO: 168); QSPTSNHSPTSCPPIC (SEQ ID NO:169); IIFLFILLLCLIFLLVLLD (SEQ ID NO: 170); CTTPAQGNSMFPSC (SEQ ID NO:171); CTKPTDGN (SEQ ID NO: 172); WASVRFSW (SEQ ID NO: 173); LLPIFFCLW(SEQ ID NO: 174); MDIDPYKEFGATVELLSFLP (SEQ ID NO: 175); FLPSDFFPSV (SEQID NO: 176); RDLLDTASALYREALESPEH (SEQ ID NO: 177); PHHTALRQAILCWGELMTLA(SEQ ID NO: 178); GRETVIEYLVSFGVW (SEQ ID NO: 179); EYLVSFGVWIRTPPA (SEQID NO: 180); VSFGVWIRTPPAYRPPNAPI (SEQ ID NO: 181); TVVRRRGRSP (SEQ IDNO: 182); VGPLTVNEKRRLKLI (SEQ ID NO: 183); RHYLHTLWKAGILYK (SEQ ID NO:184); ESRLVVDFSQFSRGN (SEQ ID NO: 185); LQSLTNLLSSNLSWL (SEQ ID NO:186); SSNLSWLSLDVSAAF (SEQ ID NO: 187); LHLYSHPIILGFRKI (SEQ ID NO:188); KQCFRKLPVNRPIDW (SEQ ID NO: 189); LCQVFADATPTGWGL (SEQ ID NO:190); AANWILRGTSFVYVP (SEQ ID NO: 191); and EIRLKVFVLGGCRHK (SEQ ID NO:192), in which the a denotes D-form alanine, the (Cha) denotesL-cyclohexylalanins, and the Z denotes 6-aminohexanoic acid.

Example 52 B₁AT₁-T₂-B₂ and B₂-T₂-T₁AB₁

The peptide unit of any one of Examples 49 and 50, wherein the peptideunit is characterized in that it further includes an auxiliary part, andthe auxiliary part is linked between the first B-cell epitope and thefirst Th epitope.

Example 53 B₁-T₁AT₂-B₂

The peptide unit of any one of Examples 49 and 50, wherein the peptideunit is characterized in that it further includes an auxiliary part, andthe auxiliary part is linked between the first Th epitope and the secondTh epitope.

Example 54 B₁-T₁-T₂AB₂ and B₂AT₂-T₁-B₁

The peptide unit of any one of Examples 49 and 50, wherein the peptideunit is characterized in that it further includes an auxiliary part, andthe auxiliary part is linked between the second Th epitope and thesecond B-cell epitope.

Example 55 Limitation on A, Including Nonstandard Amino Acid

The peptide unit of any one of Examples 52 to 54, wherein the peptideunit is characterized in that the auxiliary part includes one or morenonstandard amino acids.

Example 56 Limitation on A Including Nonstandard Amino Acid

The peptide unit of any one of Examples 52 to 54, wherein the peptideunit is characterized in that the auxiliary part is selected from a, Z,aZ, Za, GSHHHHHHGSDDDKZa (SEQ ID NO: 193), and aZGSHHHHHHGSDDDK (SEQ IDNO: 194).

Example 57 Limitation on Sequence A Not Including Nonstandard Amino Acid

The peptide unit of any one of Examples 52 to 54, wherein the peptideunit is characterized in that the auxiliary part is selected from CR,HHHHHH (SEQ ID NO: 53), and RRRRRR (SEQ ID NO: 159).

Example 58 B₁A₁T₁A₂T₂-B₂ and B₂-T₂A₂T₁A₁B₁

The peptide unit of any one of Examples 49 and 50, wherein the peptideunit is characterized in that it further includes a first auxiliary partand a second auxiliary part, and the auxiliary part is linked betweenthe first B-cell epitope and the first Th epitope, and the secondauxiliary part is linked between the first Th epitope and the second Thepitope.

Example 59 B₁A₁T₁-T₂A₂B₂

The peptide unit of any one of Examples 49 and 50, wherein the peptideunit is characterized in that it further includes a first auxiliary partand a second auxiliary part, and the auxiliary part is linked betweenthe first B-cell epitope and the first Th epitope, and the secondauxiliary part is linked between the second Th epitope and the secondB-cell epitope.

Example 60 B₁-T₁A₁T₂A₂B₂ and B₂-T₂A₂T₁A₁B₁

The peptide unit of any one of Examples 49 and 50, wherein the peptideunit is characterized in that it further includes a first auxiliary partand a second auxiliary part, and the auxiliary part is linked betweenthe first Th epitope and the second Th epitope the first Th epitope andthe second Th epitope is linked between the second Th epitope and thesecond B-cell epitope.

Example 61 Limitation on A, Including Nonstandard Amino Acid

The peptide unit of any one of Examples 58 to 60, wherein the peptideunit is characterized in that the first auxiliary part and/or a secondauxiliary part include one or more nonstandard amino acids.

Example 62 Limitation on Sequence A Including Nonstandard Amino Acid

The peptide unit of any one of Examples 58 to 60, wherein the peptideunit is characterized in that the auxiliary parts including one or morenonstandard amino acids are each independently selected from a, Z, aZ,Za, GSHHHHHHGSDDDKZa (SEQ ID NO: 193), and aZGSHHHHHHGSDDDK (SEQ ID NO:194).

Example 63 Limitation on Sequence A Including Nonstandard Amino Acid

The peptide unit of any one of Examples 58 to 60, wherein the peptideunit is characterized in that the first auxiliary part and/or a secondauxiliary part are each independently selected from CR, HHHHHH (SEQ IDNO: 53), and RRRRRR (SEQ ID NO: 159).

Example 64 B₁A₁T₁A₂T₂A₃B₂

The peptide unit of any one of Examples 49 and 50, wherein the peptideunit is characterized in that it further includes a first auxiliarypart, a second auxiliary part, and a third auxiliary part; the firstauxiliary part is linked between the first B-cell epitope and the firstTh epitope; the second auxiliary part is linked between the first Thepitope and the second Th epitope; and the third auxiliary part islinked between the second Th epitope and the second B-cell epitope.

Example 65 Limitation on A, Including Nonstandard Amino Acid

The peptide unit of Example 64, wherein the peptide unit ischaracterized in that the first auxiliary part, a second auxiliary part,and/or a third auxiliary part include one or more nonstandard aminoacids.

Example 66 Limitation on Sequence A Including Nonstandard Amino Acid

The peptide unit of Example 65, wherein the peptide unit ischaracterized in that the auxiliary parts including one or morenonstandard amino acids are each independently selected from a, Z, aZ,Za, GSHHHHHHGSDDDKZa (SEQ ID NO: 193), and aZGSHHHHHHGSDDDK (SEQ ID NO:194).

Example 67 Limitation on Sequence A Not Including Nonstandard Amino Acid

The peptide unit of Example 64, wherein the peptide unit ischaracterized in that the first auxiliary part, the second auxiliarypart, and/or the third auxiliary part are each independently selectedfrom CR, HHHHHH (SEQ ID NO: 53), and RRRRRR (SEQ ID NO: 159).

Peptide Unit 7—Unit and Additional Auxiliary Part

Example 68 AU and UA

a peptide unit including the following: a peptide unit of any one ofExamples 1 to 67; and an additional auxiliary part.

Example 69 Limitation on A, Including Nonstandard Amino Acid

The peptide unit of Example 67, wherein the peptide unit ischaracterized in that the additional auxiliary part includes one or morenonstandard amino acids.

Example 70 Limitation on Sequence A Including Nonstandard Amino Acid

The peptide unit of Example 69, wherein the peptide unit ischaracterized in that the additional auxiliary part is selected from a,Z, aZ, Za, GSHHHHHHGSDDDKZa (SEQ ID NO: 193), and aZGSHHHHHHGSDDDK (SEQID NO: 194).

Example 71 Limitation on Sequence A Not Including Nonstandard Amino Acid

The peptide unit of Example 68, wherein the peptide unit ischaracterized in that the additional auxiliary part is selected from CR,HHHHHH (SEQ ID NO: 53), and RRRRRR (SEQ ID NO: 159).

Example 72 AUA

a peptide unit including the following: a peptide unit of any one ofExamples 1 to 67; and a first additional auxiliary part and a secondadditional auxiliary part. In particular, the peptide unit is linkedbetween the first additional auxiliary part and the second additionalauxiliary part.

Example 73 Limitation on A, Including Nonstandard Amino Acid

The peptide unit of Example 72, wherein the peptide unit ischaracterized in that the first additional auxiliary part and/or thesecond additional auxiliary part include one or more nonstandard aminoacids.

Example 74 Limitation on Sequence A Including Nonstandard Amino Acid

The peptide unit of Example 73, wherein the peptide unit ischaracterized in that the additional auxiliary parts including one ormore nonstandard amino acids are each independently selected from a, Z,aZ, Za, GSHHHHHHGSDDDKZa (SEQ ID NO: 193), and aZGSHHHHHHGSDDDK (SEQ IDNO: 194).

Example 75 Limitation on Sequence A Not Including Nonstandard Amino Acid

The peptide unit of Example 72, wherein the peptide unit ischaracterized in that the first additional auxiliary part and/or and thesecond additional auxiliary part are each independently selected fromCR, HHHHHH (SEQ ID NO: 53), and RRRRRR (SEQ ID NO: 159).

Peptide Unit 8—Limitation on Unit Sequence

Example 76 Limitation on Sequence of Unit-A

The peptide unit of any one of Examples 1 to 8 and 68 to 75, wherein thepeptide unit is selected from the following:RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZ (SEQ ID NO: 56);ZaK(Cha)VAAWTLKAAaZRNVPPIFNDVYWIAF (SEQ ID NO: 57);CRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZ (SEQ ID NO: 58);ZaK(Cha)VAAWTLKAAaZCRFRGLISLSQVYLS (SEQ ID NO: 59);KTTKQSFDLSVKAQYKKNKHZaK(Cha)VAAWTLKAAaZ (SEQ ID NO: 60);ZaK(Cha)VAAWTLKAAaZKTTKQSFDLSVKAQYKKNKH (SEQ ID NO: 61);RNVPPIFNDVYWIAFK(Cha)VAAWTLKAA (SEQ ID NO: 62);K(Cha)VAAWTLKAARNVPPIFNDVYWIAF (SEQ ID NO: 63);RNVPPIFNDVYK(Cha)VAAWTLKAA (SEQ ID NO: 64);

PIFNDVYWIAFK(Cha)VAAWTLKAA (SEQ ID NO: 65); PPIFNDVYWK(Cha)VAAWTLKAA(SEQ ID NO: 66); RNVPPIFNDVYWIAFK(Cha)VAAWTLKAAHHHHHH (SEQ ID NO: 67);RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZGSHHHHHHGSDDDDK (SEQ ID NO: 68);GSHHHHHHGSDDDDKZaK(Cha)VAAWTLKAAaZRNVPPIFNDVYWIAF (SEQ ID NO: 69);RNVPPIFNDVYWIAFGSHHHHHHGSDDDDKZaK(Cha)VAAWTLKAAaZ (SEQ ID NO: 70);GSHHHHHHGSDDDDKZaK(Cha)VAAWTLKAAaZCRFRGLISLSQVYLS (SEQ ID NO: 71);GSHHHHHHGSDDDDKCRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZ (SEQ ID NO: 72);CRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZGSHHHHHHGSDDDDK (SEQ ID NO: 73);GSHHHHHHGSDDDDKZaK(Cha)VAAWTLKAAaZKTTKQSFDLSVKAQYKKNKH (SEQ ID NO: 74);GSHHHHHHGSDDDDKKTTKQSFDLSVKAQYKKNKHZaK(Cha)VAAWTLKAAaZ (SEQ ID NO: 75);KTTKQSFDLSVKAQYKKNKHZaK(Cha)VAAWTLKAAaZGSHHHHHHGSDDDDK (SEQ ID NO: 76);MRGSHHHHHHGSDDDDKIVDRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZ (SEQ ID NO: 77);MRGSHHHHHHGSDDDDKIVDGSHHHHHHGSDDDDKRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZ(SEQ ID NO: 78);MRGSHHHHHHGSDDDDKIVDRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZGS HHHHHHGSDDDDK(SEQ ID NO: 79); RNVPPIFNDVYWIAFILMQYIKANSKFIGI (SEQ ID NO: 80);RNVPPIFNDVYWIAFILMQYIKANSKFIGIPMGLPQSIALSSLMVAQ (SEQ ID NO: 81);CRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZC (SEQ ID NO: 82);RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAACR (SEQ ID NO: 161);RNVPPIFNDVYWIAFXXKXVAAWTLKAAXXGSHHHHHHGSDDDDK (SEQ ID NO: 199);GSHHHHHHGSDDDDKXXKXVAAWTLKAAXXRNVPPIFNDVYWIAF (SEQ ID NO: 200);RNVPPIFNDVYWIAFXXKXVAAWTLKAAXX (SEQ ID NO: 204);RNVPPIFNDVYWIAFKXVAAWTLKAA (SEQ ID NO: 205);RNVPPIFNDVYWIAFKXVAAWTLKAAHHHHHH (SEQ ID NO: 206);RNVPPIFNDVYWIAFXXKXVAAWTLKAACR (SEQ ID NO: 208);RNVPPIFNDVYWIAFXXKFVAAWTLKAAXX (SEQ ID NO: 210);RNVPPIFNDVYWIAFXXKFVAAWTLKAACR (SEQ ID NO: 212); RNVPPIFNDVYWIAFCTKPTDGN(SEQ ID NO: 213); RNVPPIFNDVYWIAFLLPIFFCLW (SEQ ID NO: 214);RNVPPIFNDVYWIAFFLPSDFFPSV (SEQ ID NO: 215);RNVPPIFNDVYWIAFILMQYIKANSKFIGIHHHHHH (SEQ ID NO: 219); andRNVPPIFNDVYWIAFMDIDPYKEFGATVELLSFLPHHHHHH (SEQ ID NO: 220), in which thea denotes D-form alanine, the Z denotes 6-aminohexanoic acid, the (Cha)denotes L-cyclohexylalanine, and the X denotes any standard amino acid.

Example 77 Limitation on Sequence of Unit-D

The peptide unit of any one of Examples 1 to 6, 14 to 15, and 68 to 75,wherein the peptide unit is selected from the following: RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZ ILMQYIKANSKFIGI (SEQ ID NO: 123); CRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZ ILMQYIKANSKFIGI (SEQ ID NO: 124);KTTKQSFDLSVKAQYKKNKH ZaK(Cha)VAAWTLKAAaZ ILMQYIKANSKFIGI (SEQ ID NO:125); ILMQYIKANSKFIGI ZaK(Cha)VAAWTLKAAaZ RNVPPIFNDVYWIAF (SEQ ID NO:126); ILMQYIKANSKFIGI ZaK(Cha)VAAWTLKAAaZ CRFRGLISLSQVYLS (SEQ ID NO:127); ILMQYIKANSKFIGI ZaK(Cha)VAAWTLKAAaZ KTTKQSFDLSVKAQYKKNKH (SEQ IDNO: 128); PIFNDVYWIAF K(Cha)VAAWTLKAA K(Cha)VAAWTLKAA (SEQ ID NO: 129);PPIFNDVYW K(Cha)VAAWTLKAA K(Cha)VAAWTLKAA (SEQ ID NO: 130);MRGSHHHHHHGSDDDDKIVDRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZIL MQYIKANSKFIGI(SEQ ID NO: 131); MRGSHHHHHHGSDDDDKIVDILMQYIKANSKFIGIZaK(Cha)VAAWTLKAAaZ(SEQ ID NO: 132);MRGSHHHHHHGSDDDDKIVDRNVPPIFNDVYWIAFGGGGSGGGGGGSSZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGI (SEQ ID NO: 133);MRGSHHHHHHGSDDDDKIVDRNVPPIFNDVYWIAFGGGGSGGGGGGSSILMQYIKANSKFIGIPMGLPQSIALSSLMVAQGGGGSGGGGGGSSILMQYIKANSKFIGIPM GLPQSIALSSLMVAQ(SEQ ID NO: 134); RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZK(Cha)LAAFTIRAAaZ(SEQ ID NO: 135); andCRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGIC (SEQ ID NO: 136), inwhich the a denotes D-form alanine, the Z denotes 6-aminohexanoic acid,and (Cha) denotes L-cyclohexylalanine.

Example 78 Limitation on Sequence of Unit-B

The peptide unit of any one of Examples 1 to 6, 26 to 27, and 68 to 75,wherein the peptide unit is selected from the group consisting of thefollowing: RNVPPIFNDVYWIAFRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZ (SEQ ID NO:83), RNVPPIFNDVYWIAFCRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZ (P5: SEQ ID NO:84), RNVPPIFNDVYWIAFKTTKQSFDLSVKAQYKKNKHZaK(Cha)VAAWTLKAAaZ (SEQ ID NO:85), CRFRGLISLSQVYLSRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZ (SEQ ID NO: 86),CRFRGLISLSQVYLSCRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZ (SEQ ID NO: 87),CRFRGLISLSQVYLSKTTKQSFDLSVKAQYKKNKHZaK(Cha)VAAWTLKAAaZ (SEQ ID NO: 88),KTTKQSFDLSVKAQYKKNKHRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZ (SEQ ID NO: 89),KTTKQSFDLSVKAQYKKNKHCRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZ (SEQ ID NO: 90),KTTKQSFDLSVKAQYKKNKHKTTKQSFDLSVKAQYKKNKHZaK(Cha)VAAWTLK AAaZ (SEQ ID NO:91), RNVPPIFNDVYWIAFCRFRGLISLSQVYLSK(Cha)VAAWTLKAA (SEQ ID NO: 92),PIFNDVYWIAFGLISLSQVYLSK(Cha)VAAWTLKAA (SEQ ID NO: 93),RNVPPIFNDVYCRFRGLISLSQK(Cha)VAAWTLKAA (SEQ ID NO: 94),PIFNDVYWIAFCRFRGLISLSQK(Cha)VAAWTLKAA (SEQ ID NO: 95),PPIFNDVYWRGLISLSQVK(Cha)VAAWTLKAA (SEQ ID NO: 96),RNVPPIFNDVYWIAFCRFRGLISLSQVYLSK(Cha)VAAWTLKAAHHHHHH (SEQ ID NO: 97),MRGSHHHHHHGSDDDDKIVDRNVPPIFNDVYWIAFCRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZ(SEQ ID NO: 98),MRGSHHHHHHGSDDDDKIVDRNVPPIFNDVYWIAFCRFRGLISLSQVYLSZaK(Cha)VAAWTLKAA (SEQID NO: 99), RNVPPIFNDVYWIAFCRFRGLISLSQVYLSZaK(Cha)VAAWTLKAA (SEQ ID NO:100), MRGSHHHHHHGSDDDDKIVD RNVPPIFNDVYWIAF GGGGSGGGGGGSSRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAA (SEQ ID NO: 101), RNVPPIFNDVYWIAFGGGGSGGGGGGSS RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAA (SEQ ID NO: 102),RNVPPIFNDVYWIAF GGGGSGGGGGGSS RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZ (SEQ IDNO: 103), RNVPPIFNDVYWIAFRNVPPIFNDVYWIAF ILMQYIKANSKFIGI (SEQ ID NO:104), RNVPPIFNDVYWIAFRNVPPIFNDVYWIAF ILMQYIKANSKFIGIPMGLPQSIALSSLMVAQ(SEQ ID NO: 105), CRNVPPIFNDVYWIAFCRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZC(SEQ ID NO: 106), and RNVPPIFNDVYWIAFCRFRGLISLSQVYLSXXK(Cha)VAAWTLKAAXX(SEQ ID NO: 202), in which the a denotes D-form alanine, the (Cha)denotes L-cyclohexylalanins, the Z denotes 6-aminohexanoic acid, and theX denotes any standard amino acid.

Example 79 Limitation on Sequence of Unit-C

The peptide unit of any one of Examples 1 to 6, 38 to 39, and 68 to 75,wherein the peptide unit is selected from the group consisting of thefollowing: RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZRNVPPIFNDVYWIAF (SEQ ID NO:107); RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZCRFRGLISLSQVYLS (P1: SEQ ID NO:108); RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZKTTKQSFDLSVKAQYKKNKH (SEQ ID NO:109); CRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZRNVPPIFNDVYWIAF (SEQ ID NO:110); CRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZCRFRGLISLSQVYLS (SEQ ID NO:111); RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZKTTKQSFDLSVKAQYKKNKH (SEQ ID NO:112); KTTKQSFDLSVKAQYKKNKHZaK(Cha)VAAWTLKAAaZRNVPPIFNDVYWIAF (SEQ ID NO:113); KTTKQSFDLSVKAQYKKNKHZaK(Cha)VAAWTLKAAaZCRFRGLISLSQVYLS (P4: SEQ IDNO: 114); KTTKQSFDLSVKAQYKKNKHZaK(Cha)VAAWTLKAAaZKTTKQSFDLSVKAQYK KNKH(SEQ ID NO: 115); PIFNDVYWIAFK(Cha)VAAWTLKAACRFRGLISLSQ (SEQ ID NO:116); PPIFNDVYWK(Cha)VAAWTLKAARGLISLSQV (SEQ ID NO: 117);MRGSHHHHHHGSDDDDKIVD RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZRNVPPIFNDVYWIAF(SEQ ID NO: 118); MRGSHHHHHHGSDDDDKIVDRNVPPIFNDVYWIAF GGGGSGGGGGGSSILMQYIKANSKFIGIPMGLPQSIALSSLMVAQ GGGGSGGGGGGSSCRFRGLISLSQVYLS (SEQ IDNO: 119); RNVPPIFNDVYWIAFILMQYIKANSKFIGICRFRGLISLSQVYLS (SEQ ID NO:120); RNVPPIFNDVYWIAFZPKYVKQNTLKLATZCRFRGLISLSQVYLS (P5: SEQ ID NO:121); CRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZRNVPPIFNDVYWIAFC (SEQ ID NO:122); RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAACRFRGLISLSQVYLS (SEQ ID NO: 160);RNVPPIFNDVYWIAFXXKXVAAWTLKAAXXCRFRGLISLSQVYLS (SEQ ID NO: 198);KTTKQSFDLSVKAQYKKNKHXXKXVAAWTLKAAXXCRFRGLISLSQVYLS (SEQ ID NO: 201);RNVPPIFNDVYWIAFXPKYVKQNTLKLATXCRFRGLISLSQVYLS (SEQ ID NO: 203);RNVPPIFNDVYWIAFXXKXVAAWTLKAACRFRGLISLSQVYLS (SEQ ID NO: 207);RNVPPIFNDVYWIAFXXKFVAAWTLKAAXXCRFRGLISLSQVYLS (SEQ ID NO: 209);RNVPPIFNDVYWIAFXXKFVAAWTLKAACRFRGLISLSQVYLS (SEQ ID NO: 211);KTTKQSFDLSVKAQYKKNKHZaWPEANQVGAGAFGPGFaZCRFRGLISLSQVYLS (SEQ ID NO:216); KTTKQSFDLSVKAQYKKNKHZaMDIDPYKEFGATVELLSFLPaZCRFRGLISLSQV YLS (SEQID NO: 217); andKTTKQSFDLSVKAQYKKNKHZaILMQYIKANSKFIGIPMGLPQSIALSSLMVAQaZ CRFRGLISLSQVYLS(SEQ ID NO: 218), in which the a denotes D-form alanine, the (Cha)denotes L-cyclohexylalanins, the Z denotes 6-aminohexanoic acid, and theX denotes any standard amino acid.

Example 80 Limitation on Sequence of Unit-E

The peptide unit of any one of Examples 1 to 6, 49 to 50, and 68 to 75,wherein the peptide unit is selected from the group consisting of thefollowing: RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGIRNVPPIFNDVYWIAF (SEQ ID NO: 137),RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGICRFRGLISLS QVYLS (SEQID NO: 138), RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGIKTTKQSFDLSVKAQYKKNKH (SEQ ID NO: 139),CRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGIRNVPPIFNDV YWIAF (SEQID NO: 140), CRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGICRFRGLISLSQVYLS (SEQ ID NO: 141),CRFRGLISLSQVYLSZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGIKTTKQSFDLS VKAQYKKNKH(SEQ ID NO: 142),KTTKQSFDLSVKAQYKKNKHZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGIRNV PPIFNDVYWIAF(SEQ ID NO: 143),KTTKQSFDLSVKAQYKKNKHZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGICRF RGLISLSQVYLS(SEQ ID NO: 144),KTTKQSFDLSVKAQYKKNKHZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGIKTTKQSFDLSVKAQYKKNKH (SEQ ID NO: 145),MRGSHHHHHHGSDDDDKIVDRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGIRNVPPIFNDVYWIAF (SEQ ID NO: 146),MRGSHHHHHHGSDDDDKIVDRNVPPIFNDVYWIAFGGGGSGGGGGGSSILMQYI KANSKFIGIPMGLPQSIALS SLMVAQILMQYIKANSKFIGIPMGLPQSIALSSLMVAQGGGGSGGGGGGSSCRFRGLISLSQVYLS (SEQ ID NO: 147),PIFNDVYWIAFK(Cha)VAAWTLKAAK(Cha)VAAWTLKAACRFRGLISLSQ (SEQ ID NO: 148),PPIFNDVYWK(Cha)VAAWTLKAAK(Cha)VAAWTLKAARGLISLSQV (SEQ ID NO: 149), andCRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZILMQYIKANSKFIGIRNVPPIFND VYWIAFC (SEQID NO: 150), in which the a denotes D-form alanine, the (Cha) denotesL-cyclohexylalanins, and the Z denotes 6-aminohexanoic acid.

Peptide Unit 9—Formula

Example 81 Higher Concept of Unit-A Formula

The peptide unit of any one of Examples 1 to 2, wherein the peptide unitis represented by the following [Formula A] or [Formula A′]:

A₁-B-A₂-T-A₃   [Formula A]

A₁-T-A₂-B-A₃   [Formula A′]

in which the peptide unit is one which can induce a humoral immunity bybeing recognized by CD4+ T-cells and has a length of 23mer, 24mer,25mer, 26mer, 27mer, 28mer, 29mer, 30mer, 31mer, 32mer, 33mer, 34mer,35mer, 36mer, 37mer, 38mer, 39mer, 40mer, 41mer, 42mer, 43mer, 44mer,45mer, 46mer, 47mer, 48mer, 49mer, 50mer, 51mer, 52mer, 53mer, 54mer,55mer, 56mer, 57mer, 58mer, 59mer, 60mer, 61mer, 62mer, 63mer, 64mer,65mer, 66mer, 67mer, 68mer, 69mer, 70mer, or 71mer;

in which the A₁ is a first auxiliary part or absent; wherein the firstauxiliary part has a linker function, a protective function, a cyclicform forming function, a dummy function, and/or a solubility increasingfunction, and may optionally have a nonstandard amino acid;

the A₂ is a second auxiliary part or absent; wherein the secondauxiliary part has a linker function, a protective function, a cyclicform forming function, a dummy function, and/or a solubility increasingfunction, and may optionally have a nonstandard amino acid;

the A3 is a third auxiliary part or absent; wherein the third auxiliarypart has a linker function, a protective function, a cyclic form formingfunction, a dummy function, and/or a solubility increasing function, andmay optionally have a nonstandard amino acid;

the B, which is a B-cell epitope, is a fragment of apolipoprotein B-100or a mimotope of apolipoprotein B-100 and can induce an antibody whichtargets apolipoprotein B-100;

the T, which is a Th epitope, can be recognized by CD4+ T-cells and hasa length of 8mer, 9mer, 10mer, 11mer, 12mer, 13mer, 14mer, 15mer, 16mer,17mer, 18mer, 19mer, 20mer, 21mer, 22mer, 23mer, 24mer, 25mer, 26mer,27mer, 28mer, 29mer, 30mer, 31mer, or 32mer.

Example 82 Combination of Unit-A Sequences

In Example 81,

the A₁, A₂, and A₃ may be each independently selected from the groupconsisting of a, Z, aZ, Za, RN, AF, CR, LS, KT, KH, RF, DP, SV, GL, ZRNV(SEQ ID NO: 36), aZRN (SEQ ID NO: 37), IAFZ (SEQ ID NO: 38), AFZa (SEQID NO: 39), RNVP (SEQ ID NO: 40), WIAF (SEQ ID NO: 41), ZCRF (SEQ ID NO:42), aZCR (SEQ ID NO: 43), YLSZ (SEQ ID NO: 44), LSZa (SEQ ID NO: 45),CRFR (SEQ ID NO: 46), VYLS (SEQ ID NO: 47), ZKTT (SEQ ID NO: 48), aZKT(SEQ ID NO: 49), NKHZ (SEQ ID NO: 50), KHZa (SEQ ID NO: 51),GSHHHHHHGSDDDDK (SEQ ID NO: 52), HHHHHH (SEQ ID NO: 53),MRGSHHHHHHGSDDDDKIVD (SEQ ID NO: 54), GGGGSGGGGGGSS (SEQ ID

NO: 55), RRRRRR (SEQ ID NO: 159), GSHHHHHHGSDDDDKZa (SEQ ID NO: 193),and aZGSHHRHHHGSDDDDK (SEQ ID NO: 194) or may be absent, in which the adenotes D-form alanine, and the Z denotes 6-aminohexanoic acid;

the B has:

one selected from the group consisting of RNVPPIFNDVYWIAF (SEQ ID NO:6), CRFRGLISLSQVYLS (SEQ ID NO: 7), KTTKQSFDLSVKAQYKKNKH (SEQ ID NO: 8),RNVPPIFNDVY (SEQ ID NO: 9), CRFRGLISLSQ (SEQ ID NO: 10), KTTKQSFDLSVK(SEQ ID NO: 11), RNVPPIFNDVYW (SEQ ID NO: 12), CRFRGLISLSQV (SEQ ID NO:13), KTTKQSFDLSVKAQYKK (SEQ ID NO: 14), RNVPPIFNDVYWI (SEQ ID NO: 15),CRFRGLISLSQVY (SEQ ID NO: 16), KTTKQSFDLSVKAQYKKN (SEQ ID NO: 17),PIFNDVYWIAF (SEQ ID NO: 18), GLISLSQVYLS (SEQ ID NO: 19),QSFDLSVKAQYKKNKH (SEQ ID NO: 20), PPIFNDVYWIAF (SEQ ID NO: 21),RGLISLSQVYLS (SEQ ID NO: 22), KQSFDLSVKAQYKKNKH (SEQ ID NO: 23),VPPIFNDVYWIAF (SEQ ID NO: 24), FRGLISLSQVYLS (SEQ ID NO: 25),TKQSFDLSVKAQYKKNKH (SEQ ID NO: 26), NVPPIFNDVYWIA (SEQ ID NO: 27),RFRGLISLSQVYL (SEQ ID NO: 28), TKQSFDLSVKAQYKKN (SEQ ID NO: 29),VPPIFNDVYWI (SEQ ID NO: 30), FRGLISLSQVY (SEQ ID NO: 31),TKQSFDLSVKAQYKKN (SEQ ID NO: 32), PPIFNDVYW (SEQ ID NO: 33), RGLISLSQV(SEQ ID NO: 34), KQSFDLSVKAQYKK (SEQ ID NO: 35), RFRGLISLSQVYLDP (SEQ IDNO: 221), and SVCGCPVGHHDVVGL (SEQ ID NO: 222);

an epitope, which is included in any one of SEQ ID NO: 6 to SEQ ID NO:35, and SEQ ID NO: 221 to SEQ ID NO: 222; or

a sequence which matches 80% or more, 81% or more, 82% or more, 83% ormore, 84% or more, 85% or more, 86% or more, 87% or more, 88% or more,89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% ormore, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more,or 100% or more, with any one of SEQ ID NO: 6 to SEQ ID NO: 35, and SEQID NO: 221 to SEQ ID NO: 222;

in which the a denotes D-form alanine, and the Z denotes 6-aminohexanoicacid;

the T is selected from the group consisting of the following:

one selected from a group consisting of K(Cha)VAAWTLKAA (SEQ ID NO: 1),PKYVKQNTLKLAT (SEQ ID NO: 2), ILMQYIKANSKFIGI (SEQ ID NO: 3),QSIALSSLMVAQAIP (SEQ ID NO: 4), ILMQYIKANSKFIGIPMGLPQSIALSSLMVAQ (SEQ IDNO: 5), PLGFFPDHQL (SEQ ID NO: 162), WPEANQVGAGAFGPGF (SEQ ID NO: 163),MQWNSTALHQALQDP (SEQ ID NO: 164), MQWNSTTFHQTLQDPRVRGLYFPAGG (SEQ ID NO:165), FFLLTRILTI (SEQ ID NO: 166), FFLLTRILTIPQSLD (SEQ ID NO: 167),TSLNFLGGTTVCLGQ (SEQ ID NO: 168), QSPTSNHSPTSCPPIC (SEQ ID NO: 169),IIFLFILLLCLIFLLVLLD (SEQ ID NO: 170), CTTPAQGNSMFPSC (SEQ ID NO: 171),CTKPTDGN (SEQ ID NO: 172), WASVRFSW (SEQ ID NO: 173), LLPIFFCLW (SEQ IDNO: 174), MDIDPYKEFGATVELLSFLP (SEQ ID NO: 175), FLPSDFFPSV (SEQ ID NO:176), RDLLDTASALYREALESPEH (SEQ ID NO: 177), PHHTALRQAILCWGELMTLA (SEQID NO: 178), GRETVIEYLVSFGVW (SEQ ID NO: 179), EYLVSFGVWIRTPPA (SEQ IDNO: 180), VSFGVWIRTPPAYRPPNAPI (SEQ ID NO: 181), TVVRRRGRSP (SEQ ID NO:182), VGPLTVNEKRRLKLI (SEQ ID NO: 183), RHYLHTLWKAGILYK (SEQ ID NO:184), ESRLVVDFSQFSRGN (SEQ ID NO: 185), LQSLTNLLSSNLSWL (SEQ ID NO:186), SSNLSWLSLDVSAAF (SEQ ID NO: 187), LHLYSHPIILGFRKI (SEQ ID NO:188), KQCFRKLPVNRPIDW (SEQ ID NO: 189), LCQVFADATPTGWGL (SEQ ID NO:190), AANWILRGTSFVYVP (SEQ ID NO: 191), EIRLKVFVLGGCRHK (SEQ ID NO:192), KFVAAWTLKAA (SEQ ID NO: 195), KYVAAWTLKAA (SEQ ID NO: 196),DIEKKIAKMEKASSVFNVVNS (SEQ ID NO: 223), YSGPLKAEIAQRLEDV (SEQ ID NO:224), K(Cha)VKANTLKAA (SEQ ID NO: 225), K(Cha)VKANTLKAA (SEQ ID NO:226), K(Cha)VKAWTLKAA (SEQ ID NO: 227), K(Cha)VKAWTLKAA (SEQ ID NO:228), K(Cha)VWANTLKAA (SEQ ID NO: 229), K(Cha)VWANTLKAA (SEQ ID NO:230), K(Cha)VWAYTLKAA (SEQ ID NO: 231), K(Cha)VWAVTLKAA (SEQ ID NO:232), K(Cha)VYAWTLKAA (SEQ ID NO: 233), K(Cha)VYAWTLKAA (SEQ ID NO:234), R(Cha)VRANTLKAA (SEQ ID NO: 235), K(Cha)VKAHTLKAA (SEQ ID NO:236), K(Cha)VKAHTLKAA (SEQ ID NO: 237), K(Cha)VAANTLKAA (SEQ ID NO:238), K(Cha)VAANTLKAA (SEQ ID NO: 239), K(Cha)VAAYTLKAA (SEQ ID NO:240), K(Cha)VAAYTLKAA (SEQ ID NO: 241), K(Cha)VAAWTLKAA (SEQ ID NO:242), K(Cha)VAAKTLKAA (SEQ ID NO: 243), K(Cha)VAAHTLKAA (SEQ ID NO:244), K(Cha)VAAATLKAA (SEQ ID NO: 245), K(Cha)VAAWTLKAA (SEQ ID NO:246), and K(Cha)VMAATLKAA (SEQ ID NO: 247);

a sequence represented by the following [Formula I] or [Formula II];

N-Lys-X₁-X₂-Ala-Ala-X₃-Thr-X₄-X₅-Ala-Ala-C  [Formula I]

in which the X₁ is tyrosine (Tyr), phenylalanine (Phe), orL-cyclohexylalanine;

the X₂ is a hydrophobic amino acid, or leucine (Leu), or isoleucine(Ile);

the X₃ is an aromatic or cyclic amino acid, or phenylalanine (Phe),tyrosine (Tyr), or histidine (His);

the X₄ is an aliphatic long chain amino acid, or isoleucine (Ile), orvaline (Val); and

the X₅ is a charged amino acid, or arginine (Arg), leucine (Leu),aspartic acid (Asp), glutamine (Gln), or glycine (Gly);

N-Lys-X₁-Val-X₂-Ala-X₃-Thr-Leu-Lys-Ala-Ala-C  [Formula II]

in which X₁ is tyrosine (Tyr), phenylalanine (Phe), orL-cyclohexylalanine;

the X₂ is lysine (Lys), tryptophan (Trp), tyrosine (Tyr), arginine(Arg), alanine (Ala), or methionine (Met); and

the X₃ is asparagine (Asn), tryptophan (Trp), tyrosine (Tyr), valine(Val), histidine (His), lysine (Lys), or alanine (Ala); and

a sequence which matches 80% or more, 81% or more, 82% or more, 83% ormore, 84% or more, 85% or more, 86% or more, 87% or more, 88% or more,89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% ormore, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more,or 100% or more, with any one of the sequences represented by SEQ ID NO:1 to SEQ ID NO: 5, SEQ ID NO: 162 to SEQ ID NO: 192, SEQ ID NO: 195 toSEQ ID NO: 196, SEQ ID NO: 223 to SEQ ID NO: 247, and [Formula I] or[Formula II];

in which (Cha) denotes L-cyclohexylalanins, and X denotes any standardamino acid.

Example 83 Higher Concept of Unit-B Formula

The peptide unit of any one of Examples 1 and 2, wherein the peptideunit is represented by the following [Formula B] or [Formula B′]:

A₁-B₁-A₂-B₂-A₃₋T-A₄   [Formula B]

A₁-T-A₂-B₁-A₃-B₂-A₄   [Formula B′]

in which the peptide unit is characterized in that it can induce ahumoral immunity by being recognized by CD4+ T-cells and has a length of23mer, 24mer, 25mer, 26mer, 27mer, 28mer, 29mer, 30mer, 31mer, 32mer,33mer, 34mer, 35mer, 36mer, 37mer, 38mer, 39mer, 40mer, 41mer, 42mer,43mer, 44mer, 45mer, 46mer, 47mer, 48mer, 49mer, 50mer, 51mer, 52mer,53mer, 54mer, 55mer, 56mer, 57mer, 58mer, 59mer, 60mer, 61mer, 62mer,63mer, 64mer, 65mer, 66mer, 67mer, 68mer, 69mer, 70mer, or 71mer;

in which the A₁ is a first auxiliary part or absent; wherein the firstauxiliary part has a linker function, a protective function, a cyclicform forming function, a dummy function, and/or a solubility increasingfunction, and may optionally have a nonstandard amino acid;

the A₂ is a second auxiliary part or absent; wherein the secondauxiliary part has a linker function, a protective function, a cyclicform forming function, a dummy function, and/or a solubility increasingfunction, and may optionally have a nonstandard amino acid;

the A₃ is a third auxiliary part or absent; wherein the third auxiliarypart has a linker function, a protective function, a cyclic form formingfunction, a dummy function, and/or a solubility increasing function, andmay optionally have a nonstandard amino acid;

the A₄ is a fourth auxiliary part or absent; wherein the fourthauxiliary part has a linker function, a protective function, a cyclicform forming function, a dummy function, and/or a solubility increasingfunction, and may optionally have a nonstandard amino acid;

the B₁, which is a B-cell epitope, is a fragment of apolipoprotein B-100or a mimotope of apolipoprotein B-100 and can induce an antibody whichtargets apolipoprotein B-100;

the B₂, which is a B-cell epitope, is a fragment of apolipoprotein B-100or a mimotope of apolipoprotein B-100 and can induce an antibody whichtargets apolipoprotein B-100; and

the T, which is a Th epitope, can be recognized by CD4+ T-cells and hasa length of 8mer, 9mer, 10mer, 11mer, 12mer, 13mer, 14mer, 15mer, 16mer,17mer, 18mer, 19mer, 20mer, 21mer, 22mer, 23mer, 24mer, 25mer, 26mer,27mer, 28mer, 29mer, 30mer, 31mer, or 32mer.

Example 84 Combination of Unit-B Sequences

In Example 83,

the A₁, A₂, and A₃ are each independently selected from the groupconsisting of a, Z, aZ, Za, RN, AF, CR, LS, KT, KH, RF, DP, SV, GL, ZRNV(SEQ ID NO: 36), aZRN (SEQ ID NO: 37), IAFZ (SEQ ID NO: 38), AFZa (SEQID NO: 39), RNVP (SEQ ID NO: 40), WIAF (SEQ ID NO: 41), ZCRF (SEQ ID NO:42), aZCR (SEQ ID NO: 43), YLSZ (SEQ ID NO: 44), LSZa (SEQ ID NO: 45),CRFR (SEQ ID NO: 46), VYLS (SEQ ID NO: 47), ZKTT (SEQ ID NO: 48), aZKT(SEQ ID NO: 49), NKHZ (SEQ ID NO: 50), KHZa (SEQ ID NO: 51),GSHHHHHHGSDDDDK (SEQ ID NO: 52), HHHHHH (SEQ ID NO: 0 53),MRGSHHHHHHGSDDDDKIVD (SEQ ID NO: 54), GGGGSGGGGGGSS (SEQ ID

NO: 55), RRRRRR (SEQ ID NO: 159), GSHHHHHHGSDDDDKZa (SEQ ID NO: 193),and aZGSHHHHHHGSDDDDK (SEQ ID NO: 194) or is absent; in which the adenotes D-form alanine, and the Z denotes 6-aminohexanoic acid;

the B₁ and the B₂ are each independently,

selected from the group consisting of RNVPPIFNDVYWIAF (SEQ ID NO: 6),CRFRGLISLSQVYLS (SEQ ID NO: 7), KTTKQSFDLSVKAQYKKNKH (SEQ ID NO: 8),RNVPPIFNDVY (SEQ ID NO: 9), CRFRGLISLSQ (SEQ ID NO: 10), KTTKQSFDLSVK(SEQ ID NO: 11), RNVPPIFNDVYW (SEQ ID NO: 12), CRFRGLISLSQV (SEQ ID NO:13), KTTKQSFDLSVKAQYKK (SEQ ID NO: 14), RNVPPIFNDVYWI (SEQ ID NO: 15),CRFRGLISLSQVY (SEQ ID NO: 16), KTTKQSFDLSVKAQYKKN (SEQ ID NO: 17),PIFNDVYWIAF (SEQ ID NO: 18), GLISLSQVYLS (SEQ ID NO: 19),QSFDLSVKAQYKKNKH (SEQ ID NO: 20), PPIFNDVYWIAF (SEQ ID NO: 21),RGLISLSQVYLS (SEQ ID NO: 22), KQSFDLSVKAQYKKNKH (SEQ ID NO: 23),VPPIFNDVYWIAF (SEQ ID NO: 24), FRGLISLSQVYLS (SEQ ID NO: 25),TKQSFDLSVKAQYKKNKH (SEQ ID NO: 26), NVPPIFNDVYWIA (SEQ ID NO: 27),RFRGLISLSQVYL (SEQ ID NO: 28), TKQSFDLSVKAQYKKN (SEQ ID NO: 29),VPPIFNDVYWI (SEQ ID NO: 30), FRGLISLSQVY (SEQ ID NO: 31),TKQSFDLSVKAQYKKN (SEQ ID NO: 32), PPIFNDVYW (SEQ ID NO: 33), RGLISLSQV(SEQ ID NO: 34), KQSFDLSVKAQYKK (SEQ ID NO: 35), RFRGLISLSQVYLDP (SEQ IDNO: 221), and SVCGCPVGHHDVVGL (SEQ ID NO: 222);

an epitope, which is included in any one of SEQ ID NO: 6 to SEQ ID NO:35, and SEQ ID NO: 221 to SEQ ID NO: 222; or

has a sequence which matches 80% or more, 81% or more, 82% or more, 83%or more, 84% or more, 85% or more, 86% or more, 87% or more, 88% ormore, 89% or more, 90% or more, 91% or more, 92% or more, 93% or more,94% or more, 95% or more, 96% or more, 97% or more, 98% or more, 99% ormore, or 100% or more with any one of SEQ ID NO: 6 to SEQ ID NO: 35, andSEQ ID NO: 221 to SEQ ID NO: 222;

in which the a denotes D-form alanine, and the Z denotes 6-aminohexanoicacid;

the T is selected from the group consisting of the following:

one selected from a group consisting of K(Cha)VAAWTLKAA (SEQ ID NO: 1),PKYVKQNTLKLAT (SEQ ID NO: 2), ILMQYIKANSKFIGI (SEQ ID NO: 3),QSIALSSLMVAQAIP (SEQ ID NO: 4), ILMQYIKANSKFIGIPMGLPQSIALSSLMVAQ (SEQ IDNO: 5), PLGFFPDHQL (SEQ ID NO: 162), WPEANQVGAGAFGPGF (SEQ ID NO: 163),MQWNSTALHQALQDP (SEQ ID NO: 164), MQWNSTTFHQTLQDPRVRGLYFPAGG (SEQ ID NO:165), FFLLTRILTI (SEQ ID NO: 166), FFLLTRILTIPQSLD (SEQ ID NO: 167),TSLNFLGGTTVCLGQ (SEQ ID NO: 168), QSPTSNHSPTSCPPIC (SEQ ID NO: 169),IIFLFILLLCLIFLLVLLD (SEQ ID NO: 170), CTTPAQGNSMFPSC (SEQ ID NO: 171),CTKPTDGN (SEQ ID NO: 172), WASVRFSW (SEQ ID NO: 173), LLPIFFCLW (SEQ IDNO: 174), MDIDPYKEFGATVELLSFLP (SEQ ID NO: 175), FLPSDFFPSV (SEQ ID NO:176), RDLLDTASALYREALESPEH (SEQ ID NO: 177), PHHTALRQAILCWGELMTLA (SEQID NO: 178), GRETVIEYLVSFGVW (SEQ ID NO: 179), EYLVSFGVWIRTPPA (SEQ IDNO: 180), VSFGVWIRTPPAYRPPNAPI (SEQ ID NO: 181), TVVRRRGRSP (SEQ ID NO:182), VGPLTVNEKRRLKLI (SEQ ID NO: 183), RHYLHTLWKAGILYK (SEQ ID NO:184), ESRLVVDFSQFSRGN (SEQ ID NO: 185), LQSLTNLLSSNLSWL (SEQ ID NO:186), SSNLSWLSLDVSAAF (SEQ ID NO: 187), LHLYSHPIILGFRKI (SEQ ID NO:188), KQCFRKLPVNRPIDW (SEQ ID NO: 189), LCQVFADATPTGWGL (SEQ ID NO:190), AANWILRGTSFVYVP (SEQ ID NO: 191), EIRLKVFVLGGCRHK (SEQ ID NO:192), KFVAAWTLKAA (SEQ ID NO: 195), KYVAAWTLKAA (SEQ ID NO: 196),DIEKKIAKMEKASSVFNVVNS (SEQ ID NO: 223), YSGPLKAEIAQRLEDV (SEQ ID NO:224), K(Cha)VKANTLKAA (SEQ ID NO: 225), K(Cha)VKANTLKAA (SEQ ID NO:226), K(Cha)VKAWTLKAA (SEQ ID NO: 227), K(Cha)VKAWTLKAA (SEQ ID NO:228), K(Cha)VWANTLKAA (SEQ ID NO: 229), K(Cha)VWANTLKAA (SEQ ID NO:230), K(Cha)VWAYTLKAA (SEQ ID NO: 231), K(Cha)VWAVTLKAA (SEQ ID NO:232), K(Cha)VYAWTLKAA (SEQ ID NO: 233), K(Cha)VYAWTLKAA (SEQ ID NO:234), R(Cha)VRANTLKAA (SEQ ID NO: 235), K(Cha)VKAHTLKAA (SEQ ID NO:236), K(Cha)VKAHTLKAA (SEQ ID NO: 237), K(Cha)VAANTLKAA (SEQ ID NO:238), K(Cha)VAANTLKAA (SEQ ID NO: 239), K(Cha)VAAYTLKAA (SEQ ID NO:240), K(Cha)VAAYTLKAA (SEQ ID NO: 241), K(Cha)VAAWTLKAA (SEQ ID NO:242), K(Cha)VAAKTLKAA (SEQ ID NO: 243), K(Cha)VAAHTLKAA (SEQ ID NO:244), K(Cha)VAAATLKAA (SEQ ID NO: 245), K(Cha)VAAWTLKAA (SEQ ID NO:246), and K(Cha)VMAATLKAA (SEQ ID NO: 247);

a sequence represented by the following [Formula I] or [Formula II]:

N-Lys-X₁-X₂-Ala-Ala-X₃-Thr-X₄-X₅-Ala-Ala-C  [Formula I]

in which the X₁ is tyrosine (Tyr), phenylalanine (Phe), orL-cyclohexylalanine;

the X₂ is a hydrophobic amino acid, or leucine (Leu), or isoleucine(Ile);

the X₃ is an aromatic or cyclic amino acid, or phenylalanine (Phe),tyrosine (Tyr), or histidine (His);

the X₄ is an aliphatic long chain amino acid, or isoleucine (Ile), orvaline (Val); and

the X₅ is a charged amino acid, or arginine (Arg), leucine (Leu),aspartic acid (Asp), glutamine (Gln), or glycine (Gly);

N-Lys-X₁-Val-X₂-Ala-X₃-Thr-Leu-Lys-Ala-Ala-C  [Formula II]

in which X₁ is tyrosine (Tyr), phenylalanine (Phe), orL-cyclohexylalanine;

the X₂ is lysine (Lys), tryptophan (Trp), tyrosine (Tyr), arginine(Arg), alanine (Ala), or methionine (Met); and

the X₃ is asparagine (Asn), tryptophan (Trp), tyrosine (Tyr), valine(Val), histidine (His), lysine (Lys), or alanine (Ala); and

a sequence which matches 80% or more, 81% or more, 82% or more, 83% ormore, 84% or more, 85% or more, 86% or more, 87% or more, 88% or more,89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% ormore, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more,or 100% or more, with any one of the sequences represented by SEQ ID NO:1 to SEQ ID NO: 5, SEQ ID NO: 162 to SEQ ID NO: 192, SEQ ID NO: 195 toSEQ ID NO: 196, SEQ ID NO: 223 to SEQ ID NO: 247, and [Formula I] or[Formula II];

in which (Cha) denotes L-cyclohexylalanins, and X denotes any standardamino acid.

Example 85 Higher Concept of Unit-C Formula

The peptide unit of any one of Examples 1 and 2, wherein the peptideunit is represented by the following [Formula C]:

A₁-B₁-A₂-T-A₃-B₂-A₄   [Formula C]

in which the peptide unit is characterized in that it can induce ahumoral immunity by being recognized by CD4+ T-cells and has a length of23mer, 24mer, 25mer, 26mer, 27mer, 28mer, 29mer, 30mer, 31mer, 32mer,33mer, 34mer, 35mer, 36mer, 37mer, 38mer, 39mer, 40mer, 41mer, 42mer,43mer, 44mer, 45mer, 46mer, 47mer, 48mer, 49mer, 50mer, 51mer, 52mer,53mer, 54mer, 55mer, 56mer, 57mer, 58mer, 59mer, 60mer, 61mer, 62mer,63mer, 64mer, 65mer, 66mer, 67mer, 68mer, 69mer, 70mer, or 71mer;

in which the A₁ is a first auxiliary part or absent; wherein the firstauxiliary part has a linker function, a protective function, a cyclicform forming function, a dummy function, and/or a solubility increasingfunction, and may optionally have a nonstandard amino acid;

the A₂ is a second auxiliary part or absent; wherein the secondauxiliary part has a linker function, a protective function, a cyclicform forming function, a dummy function, and/or a solubility increasingfunction, and may optionally have a nonstandard amino acid;

the A₃ is a third auxiliary part or absent; wherein the third auxiliarypart has a linker function, a protective function, a cyclic form formingfunction, a dummy function, and/or a solubility increasing function, andmay optionally have a nonstandard amino acid;

the A₄ is a fourth auxiliary part or absent; wherein the fourthauxiliary part has a linker function, a protective function, a cyclicform forming function, a dummy function, and/or a solubility increasingfunction, and may optionally have a nonstandard amino acid;

the B₁, which is a B-cell epitope, is a fragment of apolipoprotein B-100or a mimotope of apolipoprotein B-100 and can induce an antibody whichtargets apolipoprotein B-100;

the B₂, which is a B-cell epitope, is a fragment of apolipoprotein B-100or a mimotope of apolipoprotein B-100 and can induce an antibody whichtargets apolipoprotein B-100; and

the T, which is a Th epitope, can be recognized by CD4+ T-cells and hasa length of 8mer, 9mer, 10mer, 11mer, 12mer, 13mer, 14mer, 15mer, 16mer,17mer, 18mer, 19mer, 20mer, 21mer, 22mer, 23mer, 24mer, 25mer, 26mer,27mer, 28mer, 29mer, 30mer, 31mer, or 32mer.

Example 86 Combination of Unit-C Sequences

In Example 85,

the A₁, A₂, A₃, and A₄ are each independently selected from the groupconsisting of a, Z, aZ, Za, RN, AF, CR, LS, KT, KH, RF, DP, SV, GL, ZRNV(SEQ ID NO: 36), aZRN (SEQ ID NO: 37), IAFZ (SEQ ID NO: 38), AFZa (SEQID NO: 39), RNVP (SEQ ID NO: 40), WIAF (SEQ ID NO: 41), ZCRF (SEQ ID NO:42), aZCR (SEQ ID NO: 43), YLSZ (SEQ ID NO: 44), LSZa (SEQ ID NO: 45),CRFR (SEQ ID NO: 46), VYLS (SEQ ID NO: 47), ZKTT (SEQ ID NO: 48), aZKT(SEQ ID NO: 49), NKHZ (SEQ ID NO: 50), KHZa (SEQ ID NO: 51),GSHHHHHHGSDDDDK (SEQ ID NO: 52), HHHHHH (SEQ ID NO: 53),MRGSHHHHHHGSDDDDKIVD (SEQ ID NO: 54), GGGGSGGGGGGSS (SEQ ID NO: 55),RRRRRR (SEQ ID NO: 159), GSHHHHHHGSDDDDKZa (SEQ ID NO: 193), andaZGSHHHHHHGSDDDDK (SEQ ID NO: 194), or is absent; in which a denotesD-form alanine, and Z denotes 6-aminohexanoic acid;

the B₁ and B₂ each independently have one selected from the groupconsisting of:

one selected from a group consisting of RNVPPIFNDVYWIAF (SEQ ID NO: 6),CRFRGLISLSQVYLS (SEQ ID NO: 7), KTTKQSFDLSVKAQYKKNKH (SEQ ID NO: 8),RNVPPIFNDVY (SEQ ID NO: 9), CRFRGLISLSQ (SEQ ID NO: 10), KTTKQSFDLSVK(SEQ ID NO: 11), RNVPPIFNDVYW (SEQ ID NO: 12), CRFRGLISLSQV (SEQ ID NO:13), KTTKQSFDLSVKAQYKK (SEQ ID NO: 14), RNVPPIFNDVYWI (SEQ ID NO: 15),CRFRGLISLSQVY (SEQ ID NO: 16), KTTKQSFDLSVKAQYKKN (SEQ ID NO: 17),PIFNDVYWIAF (SEQ ID NO: 18), GLISLSQVYLS (SEQ ID NO: 19),QSFDLSVKAQYKKNKH (SEQ ID NO: 20), PPIFNDVYWIAF (SEQ ID NO: 21),RGLISLSQVYLS (SEQ ID NO: 22), KQSFDLSVKAQYKKNKH (SEQ ID NO: 23),VPPIFNDVYWIAF (SEQ ID NO: 24), FRGLISLSQVYLS (SEQ ID NO: 25),TKQSFDLSVKAQYKKNKH (SEQ ID NO: 26), NVPPIFNDVYWIA (SEQ ID NO: 27),RFRGLISLSQVYL (SEQ ID NO: 28), TKQSFDLSVKAQYKKN (SEQ ID NO: 29),VPPIFNDVYWI (SEQ ID NO: 30), FRGLISLSQVY (SEQ ID NO: 31),TKQSFDLSVKAQYKKN (SEQ ID NO: 32), PPIFNDVYW (SEQ ID NO: 33), RGLISLSQV(SEQ ID NO: 34), KQSFDLSVKAQYKK (SEQ ID NO: 35), RFRGLISLSQVYLDP (SEQ IDNO: 221), and SVCGCPVGHHDVVGL (SEQ ID NO: 222);

an epitope, which is included in any one of SEQ ID NO: 6 to SEQ ID NO:35, and SEQ ID NO: 221 to SEQ ID NO: 222; or

a sequence which matches 80% or more, 81% or more, 82% or more, 83% ormore, 84% or more, 85% or more, 86% or more, 87% or more, 88% or more,89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% ormore, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more,or 100% or more, with any one of the sequences represented by SEQ ID NO:6 to SEQ ID NO: 35, and SEQ ID NO: 221 to SEQ ID NO: 222;

in which “a” denotes D-form alanine, and “Z” denotes 6-aminohexanoicacid;

the T is selected from the group consisting of the following:

one selected from a group consisting of K(Cha)VAAWTLKAA (SEQ ID NO: 1),PKYVKQNTLKLAT (SEQ ID NO: 2), ILMQYIKANSKFIGI (SEQ ID NO: 3),QSIALSSLMVAQAIP (SEQ ID NO: 4), ILMQYIKANSKFIGIPMGLPQSIALSSLMVAQ (SEQ IDNO: 5), PLGFFPDHQL (SEQ ID NO: 162), WPEANQVGAGAFGPGF (SEQ ID NO: 163),MQWNSTALHQALQDP (SEQ ID NO: 164), MQWNSTTFHQTLQDPRVRGLYFPAGG (SEQ ID NO:165), FFLLTRILTI (SEQ ID NO: 166), FFLLTRILTIPQSLD (SEQ ID NO: 167),TSLNFLGGTTVCLGQ (SEQ ID NO: 168), QSPTSNHSPTSCPPIC (SEQ ID NO: 169),IIFLFILLLCLIFLLVLLD (SEQ ID NO: 170), CTTPAQGNSMFPSC (SEQ ID NO: 171),CTKPTDGN (SEQ ID NO: 172), WASVRFSW (SEQ ID NO: 173), LLPIFFCLW (SEQ IDNO: 174), MDIDPYKEFGATVELLSFLP (SEQ ID NO: 175), FLPSDFFPSV (SEQ ID NO:176), RDLLDTASALYREALESPEH (SEQ ID NO: 177), PHHTALRQAILCWGELMTLA (SEQID NO: 178), GRETVIEYLVSFGVW (SEQ ID NO: 179), EYLVSFGVWIRTPPA (SEQ IDNO: 180), VSFGVWIRTPPAYRPPNAPI (SEQ ID NO: 181), TVVRRRGRSP (SEQ ID NO:182), VGPLTVNEKRRLKLI (SEQ ID NO: 183), RHYLHTLWKAGILYK (SEQ ID NO:184), ESRLVVDFSQFSRGN (SEQ ID NO: 185), LQSLTNLLSSNLSWL (SEQ ID NO:186), SSNLSWLSLDVSAAF (SEQ ID NO: 187), LHLYSHPIILGFRKI (SEQ ID NO:188), KQCFRKLPVNRPIDW (SEQ ID NO: 189), LCQVFADATPTGWGL (SEQ ID NO:190), AANWILRGTSFVYVP (SEQ ID NO: 191), EIRLKVFVLGGCRHK (SEQ ID NO:192), KFVAAWTLKAA (SEQ ID NO: 195), KYVAAWTLKAA (SEQ ID NO: 196),DIEKKIAKMEKASSVFNVVNS (SEQ ID NO: 223), YSGPLKAEIAQRLEDV (SEQ ID NO:224), K(Cha)VKANTLKAA (SEQ ID NO: 225), K(Cha)VKANTLKAA (SEQ ID NO:226), K(Cha)VKAWTLKAA (SEQ ID NO: 227), K(Cha)VKAWTLKAA (SEQ ID NO:228), K(Cha)VWANTLKAA (SEQ ID NO: 229), K(Cha)VWANTLKAA (SEQ ID NO:230), K(Cha)VWAYTLKAA (SEQ ID NO: 231), K(Cha)VWAVTLKAA (SEQ ID NO:232), K(Cha)VYAWTLKAA (SEQ ID NO: 233), K(Cha)VYAWTLKAA (SEQ ID NO:234), R(Cha)VRANTLKAA (SEQ ID NO: 235), K(Cha)VKAHTLKAA (SEQ ID NO:236), K(Cha)VKAHTLKAA (SEQ ID NO: 237), K(Cha)VAANTLKAA (SEQ ID NO:238), K(Cha)VAANTLKAA (SEQ ID NO: 239), K(Cha)VAAYTLKAA (SEQ ID NO:240), K(Cha)VAAYTLKAA (SEQ ID NO: 241), K(Cha)VAAWTLKAA (SEQ ID NO:242), K(Cha)VAAKTLKAA (SEQ ID NO: 243), K(Cha)VAAHTLKAA (SEQ ID NO:244), K(Cha)VAAATLKAA (SEQ ID NO: 245), K(Cha)VAAWTLKAA (SEQ ID NO:246), and K(Cha)VMAATLKAA (SEQ ID NO: 247);

a sequence represented by the following [Formula I] or [Formula II]:

N-Lys-X₁-X₂-Ala-Ala-X₃-Thr-X₄-X₅-Ala-Ala-C  [Formula I]

in which the X₁ is tyrosine (Tyr), phenylalanine (Phe), orL-cyclohexylalanine;

the X₂ is a hydrophobic amino acid, or leucine (Leu), or isoleucine(Ile);

the X₃ is an aromatic or cyclic amino acid, or phenylalanine (Phe),tyrosine (Tyr), or histidine (His);

the X₄ is an aliphatic long chain amino acid, or isoleucine (Ile), orvaline (Val); and

the X₅ is a charged amino acid, or arginine (Arg), leucine (Leu),aspartic acid (Asp), glutamine (Gln), or glycine (Gly);

N-Lys-X₁-Val-X₂-Ala-X₃-Thr-Leu-Lys-Ala-Ala-C  [Formula II]

in which X₁ is tyrosine (Tyr), phenylalanine (Phe), orL-cyclohexylalanine;

the X2 is lysine (Lys), tryptophan (Trp), tyrosine (Tyr), arginine(Arg), alanine (Ala), or methionine (Met); and

the X₃ is asparagine (Asn), tryptophan (Trp), tyrosine (Tyr), valine(Val), histidine (His), lysine (Lys), or alanine (Ala); and

a sequence which matches 80% or more, 81% or more, 82% or more, 83% ormore, 84% or more, 85% or more, 86% or more, 87% or more, 88% or more,89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% ormore, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more,or 100% or more, with any one of the sequences represented by SEQ ID NO:1 to SEQ ID NO: 5, SEQ ID NO: 162 to SEQ ID NO: 192, SEQ ID NO: 195 toSEQ ID NO: 196, SEQ ID NO: 223 to SEQ ID NO: 247, and [Formula I] or[Formula II];

in which (Cha) denotes L-cyclohexylalanins, and X denotes any standardamino acid.

Example 87 Higher Concept of Unit-D Formula

The peptide unit of any one of Examples 1 and 2, wherein the peptideunit is represented by the following [Formula I] or [Formula II]:

A₁-B-A₂-T₁-A₃-T₂-A₄   [Formula D]

A₁-T₁-A₂-T₂-A₃-B-A₄   [Formula D′]

in which the peptide unit is characterized in that it can induce ahumoral immunity by being recognized by CD4+ T-cells and has a length of23mer, 24mer, 25mer, 26mer, 27mer, 28mer, 29mer, 30mer, 31mer, 32mer,33mer, 34mer, 35mer, 36mer, 37mer, 38mer, 39mer, 40mer, 41mer, 42mer,43mer, 44mer, 45mer, 46mer, 47mer, 48mer, 49mer, 50mer, 51mer, 52mer,53mer, 54mer, 55mer, 56mer, 57mer, 58mer, 59mer, 60mer, 61mer, 62mer,63mer, 64mer, 65mer, 66mer, 67mer, 68mer, 69mer, 70mer, or 71mer;

in which the A₁ is a first auxiliary part or absent; wherein the firstauxiliary part has a linker function, a protective function, a cyclicform forming function, a dummy function, and/or a solubility increasingfunction, and may optionally have a nonstandard amino acid;

the A₂ is a second auxiliary part or absent; wherein the secondauxiliary part has a linker function, a protective function, a cyclicform forming function, a dummy function, and/or a solubility increasingfunction, and optionally may have a nonstandard amino acid;

the A₃ is a third auxiliary part or absent; wherein the third auxiliarypart has a linker function, a protective function, a cyclic form formingfunction, a dummy function, and/or a solubility increasing function, andmay optionally have a nonstandard amino acid;

the A₄ is a fourth auxiliary part or absent; wherein the fourthauxiliary part has a linker function, a protective function, a cyclicform forming function, a dummy function, and/or a solubility increasingfunction, and may optionally have a nonstandard amino acid;

the B, which is a B-cell epitope, is a fragment of apolipoprotein B-100or a mimotope of apolipoprotein B-100 and can induce an antibody whichtargets apolipoprotein B-100;

the T₁, which is a Th epitope, can be recognized by CD4+ T-cells and hasa length of 8mer, 9mer, 10mer, 11mer, 12mer, 13mer, 14mer, 15mer, 16mer,17mer, 18mer, 19mer, 20mer, 21mer, 22mer, 23mer, 24mer, 25mer, 26mer,27mer, 28mer, 29mer, 30mer, 31mer, or 32mer; and

the T₂, which is a Th epitope, can be recognized by CD4+ T-cells and hasa length of 8mer, 9mer, 10mer, 11mer, 12mer, 13mer, 14mer, 15mer, 16mer,17mer, 18mer, 19mer, 20mer, 21mer, 22mer, 23mer, 24mer, 25mer, 26mer,27mer, 28mer, 29mer, 30mer, 31mer, or 32mer.

Example 88 Combination of Unit-D Sequences

In Example 87,

the A₁, A₂, A₃, and A₄ are each independently selected from the groupconsisting of a, Z, aZ, Za, RN, AF, CR, LS, KT, KH, RF, DP, SV, GL, ZRNV(SEQ ID NO: 36), aZRN (SEQ ID NO: 37), IAFZ (SEQ ID NO: 38), AFZa (SEQID NO: 39), RNVP (SEQ ID NO: 40), WIAF (SEQ ID NO: 41), ZCRF (SEQ ID NO:42), aZCR (SEQ ID NO: 43), YLSZ (SEQ ID NO: 44), LSZa (SEQ ID NO: 45),CRFR (SEQ ID NO: 46), VYLS (SEQ ID NO: 47), ZKTT (SEQ ID NO: 48), aZKT(SEQ ID NO: 49), NKHZ (SEQ ID NO: 50), KHZa (SEQ ID NO: 51),GSHHHHHHGSDDDDK (SEQ ID NO: 52), HHHHHH (SEQ ID NO: 53),MRGSHHHHHHGSDDDDKIVD (SEQ ID NO: 54), GGGGSGGGGGGSS (SEQ ID NO: 55),RRRRRR (SEQ ID NO: 159), GSHHHHHHGSDDDDKZa (SEQ ID NO: 193), andaZGSHHHHHHGSDDDDK (SEQ ID NO: 194)), or is absent; in which a denotesD-form alanine, and Z denotes 6-aminohexanoic acid;

the B has:

one selected from the group consisting of RNVPPIFNDVYWIAF (SEQ ID NO:6), CRFRGLISLSQVYLS (SEQ ID NO: 7), KTTKQSFDLSVKAQYKKNKH (SEQ ID NO: 8),RNVPPIFNDVY (SEQ ID NO: 9), CRFRGLISLSQ (SEQ ID NO: 10), KTTKQSFDLSVK(SEQ ID NO: 11), RNVPPIFNDVYW (SEQ ID NO: 12), CRFRGLISLSQV (SEQ ID NO:13), KTTKQSFDLSVKAQYKK (SEQ ID NO: 14), RNVPPIFNDVYWI (SEQ ID NO: 15),CRFRGLISLSQVY (SEQ ID NO: 16), KTTKQSFDLSVKAQYKKN (SEQ ID NO: 17),PIFNDVYWIAF (SEQ ID NO: 18), GLISLSQVYLS (SEQ ID NO: 19),QSFDLSVKAQYKKNKH (SEQ ID NO: 20), PPIFNDVYWIAF (SEQ ID NO: 21),RGLISLSQVYLS (SEQ ID NO: 22), KQSFDLSVKAQYKKNKH (SEQ ID NO: 23),VPPIFNDVYWIAF (SEQ ID NO: 24), FRGLISLSQVYLS (SEQ ID NO: 25),TKQSFDLSVKAQYKKNKH (SEQ ID NO: 26), NVPPIFNDVYWIA (SEQ ID NO: 27),RFRGLISLSQVYL (SEQ ID NO: 28), TKQSFDLSVKAQYKKN (SEQ ID NO: 29),VPPIFNDVYWI (SEQ ID NO: 30), FRGLISLSQVY (SEQ ID NO: 31),TKQSFDLSVKAQYKKN (SEQ ID NO: 32), PPIFNDVYW (SEQ ID NO: 33), RGLISLSQV(SEQ ID NO: 34), KQSFDLSVKAQYKK (SEQ ID NO: 35), RFRGLISLSQVYLDP (SEQ IDNO: 221), and SVCGCPVGHHDVVGL (SEQ ID NO: 222);

an epitope, which is included in any one of SEQ ID NO: 6 to SEQ ID NO:35, and SEQ ID NO: 221 to SEQ ID NO: 222; or

a sequence which matches 80% or more, 81% or more, 82% or more, 83% ormore, 84% or more, 85% or more, 86% or more, 87% or more, 88% or more,89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% ormore, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more,or 100% or more, with any one of the sequences represented by SEQ ID NO:6 to SEQ ID NO: 35, and SEQ ID NO: 221 to SEQ ID NO: 222;

in which a denotes D-form alanine, and Z denotes 6-aminohexanoic acid;

the T₁ and T2 are each independently selected from the group consistingof the following:

one selected from a group consisting of K(Cha)VAAWTLKAA (SEQ ID NO: 1),PKYVKQNTLKLAT (SEQ ID NO: 2), ILMQYIKANSKFIGI (SEQ ID NO: 3),QSIALSSLMVAQAIP (SEQ ID NO: 4), ILMQYIKANSKFIGIPMGLPQSIALSSLMVAQ (SEQ IDNO: 5), PLGFFPDHQL (SEQ ID NO: 162), WPEANQVGAGAFGPGF (SEQ ID NO: 163),MQWNSTALHQALQDP (SEQ ID NO: 164), MQWNSTTFHQTLQDPRVRGLYFPAGG (SEQ ID NO:165), FFLLTRILTI (SEQ ID NO: 166), FFLLTRILTIPQSLD (SEQ ID NO: 167),TSLNFLGGTTVCLGQ (SEQ ID NO: 168), QSPTSNHSPTSCPPIC (SEQ ID NO: 169),IIFLFILLLCLIFLLVLLD (SEQ ID NO: 170), CTTPAQGNSMFPSC (SEQ ID NO: 171),CTKPTDGN (SEQ ID NO: 172), WASVRFSW (SEQ ID NO: 173), LLPIFFCLW (SEQ IDNO: 174), MDIDPYKEFGATVELLSFLP (SEQ ID NO: 175), FLPSDFFPSV (SEQ ID NO:176), RDLLDTASALYREALESPEH (SEQ ID NO: 177), PHHTALRQAILCWGELMTLA (SEQID NO: 178), GRETVIEYLVSFGVW (SEQ ID NO: 179), EYLVSFGVWIRTPPA (SEQ IDNO: 180), VSFGVWIRTPPAYRPPNAPI (SEQ ID NO: 181), TVVRRRGRSP (SEQ ID NO:182), VGPLTVNEKRRLKLI (SEQ ID NO: 183), RHYLHTLWKAGILYK (SEQ ID NO:184), ESRLVVDFSQFSRGN (SEQ ID NO: 185), LQSLTNLLSSNLSWL (SEQ ID NO:186), SSNLSWLSLDVSAAF (SEQ ID NO: 187), LHLYSHPIILGFRKI (SEQ ID NO:188), KQCFRKLPVNRPIDW (SEQ ID NO: 189), LCQVFADATPTGWGL (SEQ ID NO:190), AANWILRGTSFVYVP (SEQ ID NO: 191), EIRLKVFVLGGCRHK (SEQ ID NO:192), KFVAAWTLKAA (SEQ ID NO: 195), KYVAAWTLKAA (SEQ ID NO: 196),DIEKKIAKMEKASSVFNVVNS (SEQ ID NO: 223), YSGPLKAEIAQRLEDV (SEQ ID NO:224), K(Cha)VKANTLKAA (SEQ ID NO: 225), K(Cha)VKANTLKAA (SEQ ID NO:226), K(Cha)VKAWTLKAA (SEQ ID NO: 227), K(Cha)VKAWTLKAA (SEQ ID NO:228), K(Cha)VWANTLKAA (SEQ ID NO: 229), K(Cha)VWANTLKAA (SEQ ID NO:230), K(Cha)VWAYTLKAA (SEQ ID NO: 231), K(Cha)VWAVTLKAA (SEQ ID NO:232), K(Cha)VYAWTLKAA (SEQ ID NO: 233), K(Cha)VYAWTLKAA (SEQ ID NO:234), R(Cha)VRANTLKAA (SEQ ID NO: 235), K(Cha)VKAHTLKAA (SEQ ID NO:236), K(Cha)VKAHTLKAA (SEQ ID NO: 237), K(Cha)VAANTLKAA (SEQ ID NO:238), K(Cha)VAANTLKAA (SEQ ID NO: 239), K(Cha)VAAYTLKAA (SEQ ID NO:240), K(Cha)VAAYTLKAA (SEQ ID NO: 241), K(Cha)VAAWTLKAA (SEQ ID NO:242), K(Cha)VAAKTLKAA (SEQ ID NO: 243), K(Cha)VAAHTLKAA (SEQ ID NO:244), K(Cha)VAAATLKAA (SEQ ID NO: 245), K(Cha)VAAWTLKAA (SEQ ID NO:246), and K(Cha)VMAATLKAA (SEQ ID NO: 247);

a sequence represented by the following [Formula I] or [Formula II]:

N-Lys-X₁-X₂-Ala-Ala-X₃-Thr-X₄-X₅-Ala-Ala-C  [Formula I]

in which the X₁ is tyrosine (Tyr), phenylalanine (Phe), orL-cyclohexylalanine;

the X₂ is a hydrophobic amino acid, or leucine (Leu), or isoleucine(Ile);

the X₃ is an aromatic or cyclic amino acid, or phenylalanine (Phe),tyrosine (Tyr), or histidine (His);

the X₄ is an aliphatic long chain amino acid, or isoleucine (Ile), orvaline (Val); and

the X₅ is a charged amino acid, or arginine (Arg), leucine (Leu),aspartic acid (Asp), glutamine (Gln), or glycine (Gly);

N-Lys-X₁-Val-X₂-Ala-X₃-Thr-Leu-Lys-Ala-Ala-C:   [Formula II]

in which the X₁ is tyrosine (Tyr), phenylalanine (Phe), orL-cyclohexylalanine;

the X₂ is lysine (Lys), tryptophan (Trp), tyrosine (Tyr), arginine(Arg), alanine (Ala), or methionine (Met);

the X₃ is asparagine (Asn), tryptophan (Trp), tyrosine (Tyr), valine(Val), histidine (His), lysine (Lys), or alanine (Ala); and

a sequence which matches 80% or more, 81% or more, 82% or more, 83% ormore, 84% or more, 85% or more, 86% or more, 87% or more, 88% or more,89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% ormore, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more,or 100% or more, with any one of the sequences represented by SEQ ID NO:1 to SEQ ID NO: 5, SEQ ID NO: 162 to SEQ ID NO: 192, SEQ ID NO: 195 toSEQ ID NO: 196, SEQ ID NO: 223 to SEQ ID NO: 247, and [Formula I] or[Formula II];

in which (Cha) denotes L-cyclohexylalanins, and X denotes any standardamino acid.

Example 89 Higher Concept of Unit-E Formula

The peptide unit of any one of Examples 1 and 2, wherein the peptideunit is represented by the following [Formula E]:

A₁-B₁-A₂-T₁-A₃-T₂-A₄-B₂-A₅   [Formula E]

in which the peptide unit is one which can induce a humoral immunity bybeing recognized by CD4+ T-cells and has a length of 23mer, 24mer,25mer, 26mer, 27mer, 28mer, 29mer, 30mer, 31mer, 32mer, 33mer, 34mer,35mer, 36mer, 37mer, 38mer, 39mer, 40mer, 41mer, 42mer, 43mer, 44mer,45mer, 46mer, 47mer, 48mer, 49mer, 50mer, 51mer, 52mer, 53mer, 54mer,55mer, 56mer, 57mer, 58mer, 59mer, 60mer, 61mer, 62mer, 63mer, 64mer,65mer, 66mer, 67mer, 68mer, 69mer, 70mer, or 71mer;

in which the A₁ is a first auxiliary part or absent; wherein the firstauxiliary part has a linker function, a protective function, a cyclicform forming function, a dummy function, and/or a solubility increasingfunction, and may optionally have a nonstandard amino acid;

the A₂ is a second auxiliary part or absent; wherein the secondauxiliary part has a linker function, a protective function, a cyclicform forming function, a dummy function, and/or a solubility increasingfunction, and optionally may have a nonstandard amino acid;

the A₃ is a third auxiliary part or absent; wherein the third auxiliarypart has a linker function, a protective function, a cyclic form formingfunction, a dummy function, and/or a solubility increasing function, andmay optionally have a nonstandard amino acid;

the A₄ is a fourth auxiliary part or absent; wherein the fourthauxiliary part has a linker function, a protective function, a cyclicform forming function, a dummy function, and/or a solubility increasingfunction, and may optionally have a nonstandard amino acid;

the B₁, which is a B-cell epitope, is a fragment of apolipoprotein B-100or a mimotope of apolipoprotein B-100 and can induce an antibody whichtargets apolipoprotein B-100;

the B₂, which is a B-cell epitope, is a fragment of apolipoprotein B-100or a mimotope of apolipoprotein B-100 and can induce an antibody whichtargets apolipoprotein B-100;

the T₁, which is a Th epitope, is characterized in that it can berecognized by CD4+ T-cells and has a length of 8mer, 9mer, 10mer, 11mer,12mer, 13mer, 14mer, 15mer, 16mer, 17mer, 18mer, 19mer, 20mer, 21mer,22mer, 23mer, 24mer, 25mer, 26mer, 27mer, 28mer, 29mer, 30mer, 31mer, or32mer; and

the T₂, which is a Th epitope, is characterized in that it can berecognized by CD4+ T-cells and has a length of 8mer, 9mer, 10mer, 11mer,12mer, 13mer, 14mer, 15mer, 16mer, 17mer, 18mer, 19mer, 20mer, 21mer,22mer, 23mer, 24mer, 25mer, 26mer, 27mer, 28mer, 29mer, 30mer, 31mer, or32mer.

Example 90 Combination of Unit-E Sequences

In Example 89,

the A₁, A₂, A₃, and A₄ are each independently selected from the groupconsisting of a, Z, aZ, Za, RN, AF, CR, LS, KT, KH, RF, DP, SV, GL, ZRNV(SEQ ID NO: 36), aZRN (SEQ ID NO: 37), IAFZ (SEQ ID NO: 38), AFZa (SEQID NO: 39), RNVP (SEQ ID NO: 40), WIAF (SEQ ID NO: 41), ZCRF (SEQ ID NO:42), aZCR (SEQ ID NO: 43), YLSZ (SEQ ID NO: 44), LSZa (SEQ ID NO: 45),CRFR (SEQ ID NO: 46), VYLS (SEQ ID NO: 47), ZKTT (SEQ ID NO: 48), aZKT(SEQ ID NO: 49), NKHZ (SEQ ID NO: 50), KHZa (SEQ ID NO: 51),GSHHHHHHGSDDDDK (SEQ ID NO: 52), HHHHHH (SEQ ID NO: 53),MRGSHHHHHHGSDDDDKIVD (SEQ ID NO: 54), GGGGSGGGGGGSS (SEQ ID NO: 55),RRRRRR (SEQ ID NO: 159), GSHHHHHHGSDDDDKZa (SEQ ID NO: 193), andaZGSHHHHHHGSDDDDK (SEQ ID NO: 194), or is absent; in which a denotesD-form alanine, and Z denotes 6-aminohexanoic acid;

the B₁ and B₂ each independently have one selected from the groupconsisting of;

one selected from a group consisting of RNVPPIFNDVYWIAF (SEQ ID NO: 6),CRFRGLISLSQVYLS (SEQ ID NO: 7), KTTKQSFDLSVKAQYKKNKH (SEQ ID NO: 8),RNVPPIFNDVY (SEQ ID NO: 9), CRFRGLISLSQ (SEQ ID NO: 10), KTTKQSFDLSVK(SEQ ID NO: 11), RNVPPIFNDVYW (SEQ ID NO: 12), CRFRGLISLSQV (SEQ ID NO:13), KTTKQSFDLSVKAQYKK (SEQ ID NO: 14), RNVPPIFNDVYWI (SEQ ID NO: 15),CRFRGLISLSQVY (SEQ ID NO: 16), KTTKQSFDLSVKAQYKKN (SEQ ID NO: 17),PIFNDVYWIAF (SEQ ID NO: 18), GLISLSQVYLS (SEQ ID NO: 19),QSFDLSVKAQYKKNKH (SEQ ID NO: 20), PPIFNDVYWIAF (SEQ ID NO: 21),RGLISLSQVYLS (SEQ ID NO: 22), KQSFDLSVKAQYKKNKH (SEQ ID NO: 23),VPPIFNDVYWIAF (SEQ ID NO: 24), FRGLISLSQVYLS (SEQ ID NO: 25),TKQSFDLSVKAQYKKNKH (SEQ ID NO: 26), NVPPIFNDVYWIA (SEQ ID NO: 27),RFRGLISLSQVYL (SEQ ID NO: 28), TKQSFDLSVKAQYKKN (SEQ ID NO: 29),VPPIFNDVYWI (SEQ ID NO: 30), FRGLISLSQVY (SEQ ID NO: 31),TKQSFDLSVKAQYKKN (SEQ ID NO: 32), PPIFNDVYW (SEQ ID NO: 33), RGLISLSQV(SEQ ID NO: 34), KQSFDLSVKAQYKK (SEQ ID NO: 35), RFRGLISLSQVYLDP (SEQ IDNO: 221), and SVCGCPVGHHDVVGL (SEQ ID NO: 222);

an epitope, which is included in any one of SEQ ID NO: 6 to SEQ ID NO:35, and SEQ ID NO: 221 to SEQ ID NO: 222; or

a sequence which matches 80% or more, 81% or more, 82% or more, 83% ormore, 84% or more, 85% or more, 86% or more, 87% or more, 88% or more,89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% ormore, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more,or 100% or more, with any one of the sequences represented by SEQ ID NO:6 to SEQ ID NO: 35, and SEQ ID NO: 221 to SEQ ID NO: 222;

in which a denotes D-form alanine, and Z denotes 6-aminohexanoic acid;

the T₁ and T₂ are each independently selected from the group consistingof the following:

one selected from a group consisting of K(Cha)VAAWTLKAA (SEQ ID NO: 1),PKYVKQNTLKLAT (SEQ ID NO: 2), ILMQYIKANSKFIGI (SEQ ID NO: 3),QSIALSSLMVAQAIP (SEQ ID NO: 4), ILMQYIKANSKFIGIPMGLPQSIALSSLMVAQ (SEQ IDNO: 5), PLGFFPDHQL (SEQ ID NO: 162), WPEANQVGAGAFGPGF (SEQ ID NO: 163),MQWNSTALHQALQDP (SEQ ID NO: 164), MQWNSTTFHQTLQDPRVRGLYFPAGG (SEQ ID NO:165), FFLLTRILTI (SEQ ID NO: 166), FFLLTRILTIPQSLD (SEQ ID NO: 167),TSLNFLGGTTVCLGQ (SEQ ID NO: 168), QSPTSNHSPTSCPPIC (SEQ ID NO: 169),IIFLFILLLCLIFLLVLLD (SEQ ID NO: 170), CTTPAQGNSMFPSC (SEQ ID NO: 171),CTKPTDGN (SEQ ID NO: 172), WASVRFSW (SEQ ID NO: 173), LLPIFFCLW (SEQ IDNO: 174), MDIDPYKEFGATVELLSFLP (SEQ ID NO: 175), FLPSDFFPSV (SEQ ID NO:176), RDLLDTASALYREALESPEH (SEQ ID NO: 177), PHHTALRQAILCWGELMTLA (SEQID NO: 178), GRETVIEYLVSFGVW (SEQ ID NO: 179), EYLVSFGVWIRTPPA (SEQ IDNO: 180), VSFGVWIRTPPAYRPPNAPI (SEQ ID NO: 181), TVVRRRGRSP (SEQ ID NO:182), VGPLTVNEKRRLKLI (SEQ ID NO: 183), RHYLHTLWKAGILYK (SEQ ID NO:184), ESRLVVDFSQFSRGN (SEQ ID NO: 185), LQSLTNLLSSNLSWL (SEQ ID NO:186), SSNLSWLSLDVSAAF (SEQ ID NO: 187), LHLYSHPIILGFRKI (SEQ ID NO:188), KQCFRKLPVNRPIDW (SEQ ID NO: 189), LCQVFADATPTGWGL (SEQ ID NO:190), AANWILRGTSFVYVP (SEQ ID NO: 191), EIRLKVFVLGGCRHK (SEQ ID NO:192), KFVAAWTLKAA (SEQ ID NO: 195), KYVAAWTLKAA (SEQ ID NO: 196),DIEKKIAKMEKASSVFNVVNS (SEQ ID NO: 223), YSGPLKAEIAQRLEDV (SEQ ID NO:224), K(Cha)VKANTLKAA (SEQ ID NO: 225), K(Cha)VKANTLKAA (SEQ ID NO:226), K(Cha)VKAWTLKAA (SEQ ID NO: 227), K(Cha)VKAWTLKAA (SEQ ID NO:228), K(Cha)VWANTLKAA (SEQ ID NO: 229), K(Cha)VWANTLKAA (SEQ ID NO:230), K(Cha)VWAYTLKAA (SEQ ID NO: 231), K(Cha)VWAVTLKAA (SEQ ID NO:232), K(Cha)VYAWTLKAA (SEQ ID NO: 233), K(Cha)VYAWTLKAA (SEQ ID NO:234), R(Cha)VRANTLKAA (SEQ ID NO: 235), K(Cha)VKAHTLKAA (SEQ ID NO:236), K(Cha)VKAHTLKAA (SEQ ID NO: 237), K(Cha)VAANTLKAA (SEQ ID NO:238), K(Cha)VAANTLKAA (SEQ ID NO: 239), K(Cha)VAAYTLKAA (SEQ ID NO:240), K(Cha)VAAYTLKAA (SEQ ID NO: 241), K(Cha)VAAWTLKAA (SEQ ID NO:242), K(Cha)VAAKTLKAA (SEQ ID NO: 243), K(Cha)VAAHTLKAA (SEQ ID NO:244), K(Cha)VAAATLKAA (SEQ ID NO: 245), K(Cha)VAAWTLKAA (SEQ ID NO:246), and K(Cha)VMAATLKAA (SEQ ID NO: 247);

a sequence represented by the following [Formula I] or [Formula II]:

N-Lys-X₁-X₂-Ala-Ala-X₃-Thr-X₄-X₅-Ala-Ala-C  [Formula I]

in which the X₁ is tyrosine (Tyr), phenylalanine (Phe), orL-cyclohexylalanine;

the X₂ is a hydrophobic amino acid, or leucine (Leu), or isoleucine(Ile);

the X₃ is an aromatic or cyclic amino acid, or phenylalanine (Phe),tyrosine (Tyr), or histidine (His);

the X₄ is an aliphatic long chain amino acid, or isoleucine (Ile), orvaline (Val); and

the X₅ is a charged amino acid, or arginine (Arg), leucine (Leu),aspartic acid (Asp), glutamine (Gln), or glycine (Gly);

N-Lys-X₁-Val-X₂-Ala-X₃-Thr-Leu-Lys-Ala-Ala-C:   [Formula II]

in which X₁ is tyrosine (Tyr), phenylalanine (Phe), orL-cyclohexylalanine;

the X₂ is lysine (Lys), tryptophan (Trp), tyrosine (Tyr), arginine(Arg), alanine (Ala), or methionine (Met); and

the X₃ is asparagine (Asn), tryptophan (Trp), tyrosine (Tyr), valine(Val), histidine (His), lysine (Lys), or alanine (Ala); and

a sequence which matches 80% or more, 81% or more, 82% or more, 83% ormore, 84% or more, 85% or more, 86% or more, 87% or more, 88% or more,89% or more, 90% or more, 91% or more, 92% or more, 93% or more, 94% ormore, 95% or more, 96% or more, 97% or more, 98% or more, 99% or more,or 100% or more, with any one of the sequences represented by SEQ ID NO:1 to SEQ ID NO: 5, SEQ ID NO: 162 to SEQ ID NO: 192, SEQ ID NO: 195 toSEQ ID NO: 196, SEQ ID NO: 223 to SEQ ID NO: 247, and [Formula I] or[Formula II];

in which (Cha) denotes L-cyclohexylalanins, and X denotes any standardamino acid.

Peptide Including Peptide Unit

Example 91 Peptide Including 2 or More Peptide Units

A peptide, in which 2 or more peptide units of any one of Examples 1 to90 are linked.

Example 92 Peptide Including Two to Eight Peptide Units

A peptide, in which 2, 3, 4, 5, 6, 7, or 8 of peptide units of any oneof Examples 1 to 90 are linked.

Example 93 Concatemer Sequence

The peptide of Example 91, wherein each of the peptide units has thesame or equivalent sequence.

Example 94 String of Beads

The peptide of Example 91, wherein each of the peptide units has adifferent sequence.

Example 95 Cyclic Form

The peptide of Example 91, in which the peptide is characterized in thatit further includes an auxiliary part having a cyclic-form-formingfunction at the N-terminus and C-terminus, and the peptide forms acyclic form through the auxiliary part.

Nucleic Acid Encoding Peptide Unit and/or Peptide

Example 96 Encoding Nucleic Acid, Not Including Nonstandard Amino Acid

A nucleic acid which encodes a peptide unit of any one of Examples 1 to6 and/or a peptide of any one of Example 91 to Example 95, in which thepeptide unit and the peptide do not include a nonstandard amino acid.

Example 97 Encoding Nucleic Acid of Each Unit Formula, Not IncludingNonstandard Amino Acid

A nucleic acid, which encodes a peptide unit of any one of Example 7 toExample 10, Example 14 to Example 18, Example 22, Example 26 to Example30, Example 34, Example 38 to Example 41, Example 45, Example 49 toExample 54, Example 58 to Example 60, Example 64, Example 68, andExample 72, in which the peptide unit is characterized in that it doesnot include a nonstandard amino acid.

Example 98 Limitation on Sequence of Unit Peptide

The nucleic acid of any one of Examples 96 and 97, wherein the nucleicacid encoding a peptide unit selected from the following:RNVPPIFNDVYWIAFXXKXVAAWTLKAAXXCRFRGLISLSQVYLS (SEQ ID NO: 198);RNVPPIFNDVYWIAFXXKXVAAWTLKAAXXGSHHHHHHGSDDDDK (SEQ ID NO: 199);GSHHHHHHGSDDDDKXXKXVAAWTLKAAXXRNVPPIFNDVYWIAF (SEQ ID NO: 200);KTTKQSFDLSVKAQYKKNKHXXKXVAAWTLKAAXXCRFRGLISLSQVYLS (SEQ ID NO: 201);RNVPPIFNDVYWIAFCRFRGLISLSQVYLSXXKXVAAWTLKAAXX (SEQ ID NO: 202);RNVPPIFNDVYWIAFXPKYVKQNTLKLATXCRFRGLISLSQVYLS (SEQ ID NO: 203);RNVPPIFNDVYWIAFXXKXVAAWTLKAAXX (SEQ ID NO: 204);RNVPPIFNDVYWIAFKXVAAWTLKAA (SEQ ID NO: 205);RNVPPIFNDVYWIAFKXVAAWTLKAAHHHHHH (SEQ ID NO: 206);RNVPPIFNDVYWIAFXXKXVAAWTLKAACRFRGLISLSQVYLS (SEQ ID NO: 207);RNVPPIFNDVYWIAFXXKXVAAWTLKAACR (SEQ ID NO: 208);RNVPPIFNDVYWIAFXXKFVAAWTLKAAXXCRFRGLISLSQVYLS (SEQ ID NO: 209);RNVPPIFNDVYWIAFXXKFVAAWTLKAAXX (SEQ ID NO: 210);RNVPPIFNDVYWIAFXXKFVAAWTLKAACRFRGLISLSQVYLS (SEQ ID NO: 211); andRNVPPIFNDVYWIAFXXKFVAAWTLKAACR (SEQ ID NO: 212), in which the X denotesany standard amino acid.

Example 99 Limitation on Sequence of Unit-Encoding DNA

The nucleic acid of Example 98, a DNA which is represented by a sequenceselected from the following:

(SEQ ID NO: 248) 5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGNNNGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNNTGCCGTTTCCGTGGACTGATTTCCCTGTCCCAGGTTTATCTGTCC-3′; (SEQ ID NO: 249)5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGTTCGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNNTGCCGTTTCCGTGGACTGATTTCCCTGTCCCAGGTTTATCTGTCC-3′; (SEQ ID NO: 250)5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGTATGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNNTGCCGTTTCCGTGGACTGATTTCCCTGTCCCAGGTTTATCTGTCC-3′; (SEQ ID NO: 251)5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGNNNGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNNGGATCGCATCACCATCACCATCACGGATCCGATGATGATGACAAG-3′; (SEQ ID NO: 252)5′-ACGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGTTCGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNNGGATCGCATCACCATCACCATCACGGATCCGATGATGATGACAAG-3′; (SEQ ID NO: 253)5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGTATGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNNGGATCGCATCACCATCACCATCACGGATCCGATGATGATGACAAG-3′; (SEQ ID NO: 254)5′-GGATCGCATCACCATCACCATCACGGATCCGATGATGATGACAAGNNNNNNAAGNNNGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNNCGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTC-3′; (SEQ ID NO: 255)5′-GGATCGCATCACCATCACCATCACGGATCCGATGATGATGACAAGNNNNNNAAGTTCGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNNCGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTC-3′; (SEQ ID NO: 256)5′-GGATCGCATCACCATCACCATCACGGATCCGATGATGATGACAAGNNNNNNAAGTATGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNNCGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTC-3′; (SEQ ID NO: 257)5′-AAAACGACAAAGCAATCATTTGATTTAAGTGTAAAAGCTCAGTATNNNNNNAAGNNNGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNNTGCCGTTTCCGTGGACTGATTTCCCTGTCCCAGGTTTATCTGTCC-3′; (SEQ ID NO: 258)5′-AAAACGACAAAGCAATCATTTGATTTAAGTGTAAAAGCTCAGTATNNNNNNAAGTTCGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNNTGCCGTTTCCGTGGACTGATTTCCCTGTCCCAGGTTTATCTGTCC-3′; (SEQ ID NO: 259)5′-AAAACGACAAAGCAATCATTTGATTTAAGTGTAAAAGCTCAGTATNNNNNNAAGTATGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNNTGCCGTTTCCGTGGACTGATTTCCCTGTCCCAGGTTTATCTGTCC-3′; (SEQ ID NO: 260)5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCTGCCGTTTCCGTGGACTGATTTCCCTGTCCCAGGTTTATCTGTCCNNNNNNAAGNNNGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNN-3′; (SEQ ID NO: 261)5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCTGCCGTTTCCGTGGACTGATTTCCCTGTCCCAGGTTTATCTGTCCNNNNNNAAGTTCGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNN-3′; (SEQ ID NO: 262)5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCTGCCGTTTCCGTGGACTGATTTCCCTGTCCCAGGTTTATCTGTCCNNNNNNAAGTATGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNN-3′; (SEQ ID NO: 263)5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNCCTAAGTATGTGAAGCAGAATACACTGAAGCTGGCAACCNNNTGCCGTTTCCGTGGACTGATTTCCCTGTCCCAGGTTTATCTGTCC-3′; (SEQ ID NO: 264)5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGNNNGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNN-3′; (SEQ ID NO: 265)5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGTTCGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNN-3′; (SEQ ID NO: 266)5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGTATGTGGCAGCTTGGACCCTGAAGGCAGCANNNNNN-3′; (SEQ ID NO: 267)5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCAAGNNNGTGGCAGCTTGGACCCTGAAGGCAGCA-3′; (SEQ ID NO: 268)5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCAAGTTCGTGGCAGCTTGGACCCTGAAGGCAGCA-3′; (SEQ ID NO: 269)5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCAAGTATGTGGCAGCTTGGACCCTGAAGGCAGCA-3′; (SEQ ID NO: 270)5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCAAGNNNGTGGCAGCTTGGACCCTGAAGGCAGCACATCACCATCACCATCAC- 3′; (SEQ ID NO: 271)5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCAAGTTCGTGGCAGCTTGGACCCTGAAGGCAGCACATCACCATCACCATCAC- 3′; (SEQ ID NO: 272)5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCAAGTATGTGGCAGCTTGGACCCTGAAGGCAGCACATCACCATCACCATCAC- 3′; (SEQ ID NO: 273)5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGNNNGTGGCAGCTTGGACCCTGAAGGCAGCATGCCGTTTCCGTGGACTGATTTCCCTGTCCCAGGTTTATCTGTCC-3′; (SEQ ID NO: 274)5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGTTCGTGGCAGCTTGGACCCTGAAGGCAGCATGCCGTTTCCGTGGACTGATTTCCCTGTCCCAGGTTTATCTGTCC-3′; (SEQ ID NO: 275)5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGTATGTGGCAGCTTGGACCCTGAAGGCAGCATGCCGTTTCCGTGGACTGATTTCCCTGTCCCAGGTTTATCTGTCC-3′; (SEQ ID NO: 276)5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGNNNGTGGCAGCTTGGACCCTGAAGGCAGCATGCCGT-3′; (SEQ ID NO: 277)5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGTTCGTGGCAGCTTGGACCCTGAAGGCAGCATGCCGT-3′; and (SEQ ID NO: 278)5′-CGTAATGTTCCTCCTATCTTCAATGATGTTTATTGGATTGCATTCNNNNNNAAGTATGTGGCAGCTTGGACCCTGAAGGCAGCATGCCGT-3′.

Example 100 Limitation on Sequence of Unit-Encoding RNA

The nucleic acid of Example 98, a RNA represented by a sequence selectedfrom the following:

(SEQ ID NO: 279) 5′-CGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUCNNNNNNAAGNNNGUGGCAGCUUGGACCCUGAAGGCAGCANNNNNNUGCCGUUUCCGUGGACUGAUUUCCCUGUCCCAGGUUUAUCUGUCC-3′; (SEQ ID NO: 280)5′-CGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUCNNNNNNAAGUUCGUGGCAGCUUGGACCCUGAAGGCAGCANNNNNNUGCCGUUUCCGUGGACUGAUUUCCCUGUCCCAGGUUUAUCUGUCC-3′; (SEQ ID NO: 281)5′-CGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUCNNNNNNAAGUAUGUGGCAGCUUGGACCCUGAAGGCAGCANNNNNNUGCCGUUUCCGUGGACUGAUUUCCCUGUCCCAGGUUUAUCUGUCC-3′; (SEQ ID NO: 282)5′-CGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUCNNNNNNAAGNNNGUGGCAGCUUGGACCCUGAAGGCAGCANNNNNNGGAUCGCAUCACCAUCACCAUCACGGAUCCGAUGAUGAUGACAAG-3′; (SEQ ID NO: 283)5′-CGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUCNNNNNNAAGUUCGUGGCAGCUUGGACCCUGAAGGCAGCANNNNNNGGAUCGCAUCACCAUCACCAUCACGGAUCCGAUGAUGAUGACAAG-3′; (SEQ ID NO: 284)5′-CGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUCNNNNNNAAGUAUGUGGCAGCUUGGACCCUGAAGGCAGCANNNNNNGGAUCGCAUCACCAUCACCAUCACGGAUCCGAUGAUGAUGACAAG-3′; (SEQ ID NO: 285)5′-GGAUCGCAUCACCAUCACCAUCACGGAUCCGAUGAUGAUGACAAGNNNNNNAAGNNNGUGGCAGCUUGGACCCUGAAGGCAGCANNNNNNCGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUC-3′; (SEQ ID NO: 286)5′-GGAUCGCAUCACCAUCACCAUCACGGAUCCGAUGAUGAUGACAAGNNNNNNAAGUUCGUGGCAGCUUGGACCCUGAAGGCAGCANNNNNNCGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUC-3′; (SEQ ID NO: 287)5′-GGAUCGCAUCACCAUCACCAUCACGGAUCCGAUGAUGAUGACAAGNNNNNNAAGUAUGUGGCAGCUUGGACCCUGAAGGCAGCANNNNNNCGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUC-3′; (SEQ ID NO: 288)5′-AAAACGACAAAGCAAUCAUUUGAUUUAAGUGUAAAAGCUCAGUAUNNNNNNAAGNNNGUGGCAGCUUGGACCCUGAAGGCAGCANNNNNNUGCCGUUUCCGUGGACUGAUUUCCCUGUCCCAGGUUUAUCUGUCC-3′; (SEQ ID NO: 289)5′-AAAACGACAAAGCAAUCAUUUGAUUUAAGUGUAAAAGCUCAGUAUNNNNNNAAGUUCGUGGCAGCUUGGACCCUGAAGGCAGCANNNNNNUGCCGUUUCCGUGGACUGAUUUCCCUGUCCCAGGUUUAUCUGUCC-3′; (SEQ ID NO: 290)5′-AAAACGACAAAGCAAUCAUUUGAUUUAAGUGUAAAAGCUCAGUAUNNNNNNAAGUAUGUGGCAGCUUGGACCCUGAAGGCAGCANNNNNNUGCCGUUUCCGUGGACUGAUUUCCCUGUCCCAGGUUUAUCUGUCC-3′; (SEQ ID NO: 291)5′-CGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUCUGCCGUUUCCGUGGACUGAUUUCCCUGUCCCAGGUUUAUCUGUCCNNNNNNAAGNNNGUGGCAGCUUGGACCCUGAAGGCAGCANNNNNN-3′; (SEQ ID NO: 292)5′-CGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUCUGCCGUUUCCGUGGACUGAUUUCCCUGUCCCAGGUUUAUCUGUCCNNNNNNAAGUUCGUGGCAGCUUGGACCCUGAAGGCAGCANNNNNN-3′; (SEQ ID NO: 293)5′-CGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUCUGCCGUUUCCGUGGACUGAUUUCCCUGUCCCAGGUUUAUCUGUCCNNNNNNAAGUAUGUGGCAGCUUGGACCCUGAAGGCAGCANNNNNN-3′; (SEQ ID NO: 294)5′-CGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUCNNNCCUAAGUAUGUGAAGCAGAAUACACUGAAGCUGGCAACCNNNUGCCGUUUCCGUGGACUGAUUUCCCUGUCCCAGGUUUAUCUGUCC-3′; (SEQ ID NO: 295)5′-CGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUCNNNNNNAAGNNNGUGGCAGCUUGGACCCUGAAGGCAGCANNNNNN-3′; (SEQ ID NO: 296)5′-CGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUCNNNNNNAAGUUCGUGGCAGCUUGGACCCUGAAGGCAGCANNNNNN-3′; (SEQ ID NO: 297)5′-CGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUCNNNNNNAAGUAUGUGGCAGCUUGGACCCUGAAGGCAGCANNNNNN-3′; (SEQ ID NO: 298)5′-CGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUCAAGNNNGUGGCAGCUUGGACCCUGAAGGCAGCA-3′; (SEQ ID NO: 299)5′-CGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUCAAGUUCGUGGCAGCUUGGACCCUGAAGGCAGCA-3′; (SEQ ID NO: 300)5′-CGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUCAAGUAUGUGGCAGCUUGGACCCUGAAGGCAGCA-3′; (SEQ ID NO: 301)5′-CGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUCAAGNNNGUGGCAGCUUGGACCCUGAAGGCAGCACAUCACCAUCACCAUCAC- 3′; (SEQ ID NO: 302)5′-CGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUCAAGUUCGUGGCAGCUUGGACCCUGAAGGCAGCACAUCACCAUCACCAUCAC- 3′; (SEQ ID NO: 303)5′-CGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUCAAGUAUGUGGCAGCUUGGACCCUGAAGGCAGCACAUCACCAUCACCAUCAC- 3′; (SEQ ID NO: 304)5′-CGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUCNNNNNNAAGNNNGUGGCAGCUUGGACCCUGAAGGCAGCAUGCCGUUUCCGUGGACUGAUUUCCCUGUCCCAGGUUUAUCUGUCC-3′; (SEQ ID NO: 305)5′-CGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUCNNNNNNAAGUUCGUGGCAGCUUGGACCCUGAAGGCAGCAUGCCGUUUCCGUGGACUGAUUUCCCUGUCCCAGGUUUAUCUGUCC-3′; (SEQ ID NO: 306)5′-CGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUCNNNNNNAAGUAUGUGGCAGCUUGGACCCUGAAGGCAGCAUGCCGUUUCCGUGGACUGAUUUCCCUGUCCCAGGUUUAUCUGUCC-3′; (SEQ ID NO: 307)5′-CGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUCNNNNNNAAGNNNGUGGCAGCUUGGACCCUGAAGGCAGCAUGCCGU-3′; (SEQ ID NO: 308)5′-CGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUCNNNNNNAAGUUCGUGGCAGCUUGGACCCUGAAGGCAGCAUGCCGU-3′; and (SEQ ID NO: 309)5′-CGUAAUGUUCCUCCUAUCUUCAAUGAUGUUUAUUGGAUUGCAUUCNNNNNNAAGUAUGUGGCAGCUUGGACCCUGAAGGCAGCAUGCCGU-3′.

Example 101 Vector

A vector including any one nucleic acid of Example 96 to Example 100.

Example 102 Limitation on Vector

The vector of Example 101, wherein the vector is selected from the groupconsisting of plasmids, retroviruses, lentiviruses, adenoviruses,adeno-associated viruses, vaccinia viruses, poxviruses, and herpessimplex viruses.

Example 103 Mammalian Codon-Optimized Nucleic Acid

A nucleic acid of any one of Examples 96 to 98, wherein the nucleic acidis codon-optimized for a species selected from mammals.

Example 104 Human Codon-Optimized Nucleic Acid

A nucleic acid of any one of Examples 96 to 98, wherein the nucleic acidis human codon-optimized.

Example 105 Prokaryotic Codon-Optimized Nucleic Acid

A nucleic acid of any one of Examples 96 to 98, wherein the nucleic acidis codon-optimized for a species selected from prokaryotes.

Example 106 E. coli Codon-Optimized Nucleic Acid

A nucleic acid of any one of Examples 96 to 98, wherein the nucleic acidis E. coli codon-optimized.

Pharmaceutical Composition Including Peptide

Example 107 Pharmaceutical Composition for Immunotherapy

A pharmaceutical composition for immunotherapy including the following:

a peptide unit of any one of Examples 1 to 90, and/or a peptide of anyone of Examples 91 to 95; and

adjuvants.

Example 108 Pharmaceutical Composition for Obesity Treatment

A pharmaceutical composition for obesity treatment including thefollowing:

a peptide unit of any one of Examples 1 to 90, and/or a peptide of anyone of Examples 91 to 95; and

adjuvants.

Example 109 Limitation on Adjuvants

In any one of Examples 107 to 108, the adjuvant is water, saline,dextrose, ethanol, glycerol, sodium chloride, dextrose, mannitol,sorbitol, lactose, gelatin, albumin, aluminum hydroxide, Freund'sIncomplete Adjuvant and Complete Adjuvant (Pifco Laboratories, Detroit,Mich.), a Merck antigen adjuvant 65 (Merck and Company, Inc., Rahway,N.J.), alhydrogel (Al(OH)₃), aluminum hydroxide gel (alum), or aluminumsalts such as aluminum phosphate, AS04 series, 1VIF, squalene, MF59,QS21, calcium, iron or zinc salts, insoluble suspensions of acylatedtyrosine, acylated fructose, cationically or anionically derivedpolysaccharides, polyphosphazenes, biodegradable microspheres, Quil A,toll-like receptor (TLR) agonists, PHAD [Avanti polar lipid,Monophosphoryl Lipid A (synthetic)], monophosphoryl lipid A (MPL,monophosphoryl Lipid A), synthetic lipid A, lipid A mimics or analogues,aluminum salts, cytokines, saponins, prolactin, growth hormonedeoxycholic acid, betaglucan, polyribonucleotides, muramyl dipeptide(MDP) derivatives, CpG oligos, gram-negative bacteriallipopolysaccharide (LPS), polyphosphazene, emulsions, virosome,cochleate, poly(lactide-co-glycolide) (PLG) microparticles, poloxamerparticles, microparticles, liposomes, or appropriate combinationsthereof.

Pharmaceutical Composition Including Encoding Nucleic Acid

Example 110 Formulated Encoding Nucleic Acid

Formulated encoding nucleic acid, which is characterized in that thenucleic acid of any one of Examples 96 to 106 is formulated using aviral vector and/or non-viral vector.

Example 111 Limitation on Viral Vector

In Example 110, the encoding nucleic acid is characterized in that theviral vector is selected from the following:

retrovirus; lentivirus; adenovirus; adeno-associated virus; vacciniavirus; poxvirus; and herpes simplex virus.

Example 112 Limitation on Forms of Formulated Nucleic Acid

In Example 110, the encoding nucleic acid is characterized in that theformulated nucleic acid is selected from the following:

a naked nucleic acid; a cationic peptide-complex nucleic acid(protamine); a positively charged oil-water cationic nanoemulsionincluding a nucleic acid (cationic nanoemulsion); a nucleic acid whichis coupled with a chemically modified dendrimer and complexed withpolyethylene glycol and PEG-lipid (modified dendrimer nanoparticle); anucleic acid complexed with protamine in PEG-lipid nanoparticles(Protamine liposome); a nucleic acid which is complexed with a cationicpolymer such as polyethylenimine and PEI (cationic polymer); a nucleicacid which is complexed with a cationic polymer such as a PEI and lipidcomponent (cationic polymer liposome); a nucleic acid which is complexedwith a polysaccharide polymer such as chitosan (polysaccharideparticle); a nucleic acid which is complexed with a cationic lipidnanoparticle polymer (cationic lipid nanoparticle); a nucleic acid whichis complexed with cationic lipid and cholesterol (cationiclipid-cholesterol nanoparticle); and a nucleic acid which is complexedwith cationic lipid, cholesterol, and PEG-lipid (cationiclipid-cholesterol-PEG nanoparticle).

Example 113 Pharmaceutical Composition for Immunotherapy IncludingFormulated Nucleic Acid

A pharmaceutical composition for immunotherapy including the following:

a formulated nucleic acid of any one of Examples 100 to 112; and

adjuvants.

Example 114 Pharmaceutical Composition for Obesity Treatment IncludingFormulated Nucleic Acid

A pharmaceutical composition for obesity treatment including thefollowing:

a formulated nucleic acid of any one of Examples 100 to 112; and

adjuvants.

Example 115 Limitation on Adjuvants

A pharmaceutical composition of any one of Examples 113 and 114, whichis characterized in that the adjuvant is one or more selected from thefollowing:

lipid nanoparticles (LNPs); aluminum salts;1;2-dioleyl-3-trimethylammonium-propane chloride; MF59 (Novartis)adjuvant; CD70; CD40 ligand (CD40L); TriMix; protamine acting throughTLR7 signaling; and/or bacteria-derived monophosphoryl lipid A.

Example 116 Including Additional Ingredients

The pharmaceutical composition of any one of Examples 113 to 114,wherein the pharmaceutical composition includes one or more additionalingredients selected from the following:

lipids; salts to balance body acidity; sucrose to maintain stabilityduring repeated freezing-thawing; and vaccine stability enhancingsubstances.

Example 117 Limitation on Additional Ingredients

The pharmaceutical composition of Example 116,

wherein a pharmaceutical composition which is characterized in that thelipid is one or more selected from SM-102, PEG2000-DMG, DPSC,cholesterol, and ALC-0315;

the salt is one or more selected from sodium acetate, potassiumchloride, monobasic potassium phosphate, sodium chloride, and dibasicsodium phosphate dehydrate; and

the vaccine stability enhancing substance one or more selected fromacetic acid, an acid stabilizer (tromethamine), and ethanol.

use of peptides

Example 118 Use for Immunotherapy (First Medical Use)

Use for immunotherapy of a peptide unit of any one of Examples 1 to 90,a peptide of any one of Examples 91 to 95, a nucleic acid of any one ofExamples 96 to 106, and/or a pharmaceutical composition of any one ofExamples 107 to 109 and Examples 113 to Example 117.

Example 119 Use for Obesity Treatment (First Medical Use)

Use for obesity treatment of a peptide unit of any one of Examples 1 to90, a peptide of any one of Examples 91 to 95, a nucleic acid of any oneof Examples 96 to 106, and/or a pharmaceutical composition of any one ofExamples 107 to 109 and Examples 113 to Example 117.

Example 120 Use for Preparation of Immunotherapeutics (Second MedicalUse)

Use for preparation of immunotherapeutics of a peptide unit of any oneof Examples 1 to 90, a peptide of any one of Examples 91 to 95, anucleic acid of any one of Examples 96 to 106, and/or a pharmaceuticalcomposition of any one of Examples 107 to 109 and Examples 113 toExample 117.

Example 121 Use for Preparation of Therapeutics for Obesity Treatment(Second Medical Use)

Use for preparation of therapeutics for obesity treatment of a peptideunit of any one of Examples 1 to 90, a peptide of any one of Examples 91to 95, a nucleic acid of any one of Examples 96 to 106, and/or apharmaceutical composition of any one of Examples 107 to 109 andExamples 113 to Example 117.

Treatment Methods Using Peptides

Example 122 Immunotherapy Using Peptides

an immunotherapy including the following:

administering a peptide unit of any one of Examples 1 to 90, a peptideof any one of Examples 91 to 95, a nucleic acid of any one of Examples96 to 106, and/or a pharmaceutical composition of any one of Examples107 to 109 and Examples 113 to Example 117 into the body of a subject.

Example 123 Methods for Obesity Treatment Using Peptides

Methods for obesity treatment using the following:

administering a peptide unit of any one of Examples 1 to 90, a peptideof any one of Examples 91 to 95, a nucleic acid of any one of Examples96 to 106, and/or a pharmaceutical composition of any one of Examples107 to 109 and Examples 113 to Example 117 into the body of a subject.

Sequences Similar to Peptide Units and/or Peptides

Example 124 Sequences Similar to Peptide Units

A peptide unit which has a sequence that matches 80% or more, 81% ormore, 82% or more, 83% or more, 84% or more, 85% or more, 86% or more,87% or more, 88% or more, 89% or more, 90% or more, 91% or more, 92% ormore, 93% or more, 94% or more, 95% or more, 96% or more, 97% or more,98% or more, 99% or more, or 100% or more, with any one peptide unit ofExamples 1 to 90.

Example 125 Sequences Similar to Peptides

A peptide which has a sequence that matches 80% or more, 81% or more,82% or more, 83% or more, 84% or more, 85% or more, 86% or more, 87% ormore, 88% or more, 89% or more, 90% or more, 91% or more, 92% or more,93% or more, 94% or more, 95% or more, 96% or more, 97% or more, 98% ormore, 99% or more, or 100% or more, with any one peptide unit ofExamples 91 to 95.

EXPERIMENTAL EXAMPLES

Hereinafter, the invention provided by the present specification will bedescribed in more detail through Experimental Examples and Examples.These Examples are only for the purpose of illustrating the contentsdisclosed by the present specification, and it will be apparent to thoseskilled in the art that the scope of the contents disclosed by thepresent specification is not to be construed as being limited by theseExamples.

Experimental Example 1 Experimental Method Experimental Example 1.1Preparation of Peptides

This peptide was obtained by requesting a peptide synthesis company(Anygen, Korea, Gwangju City). The peptide of the present invention canbe synthesized using conventionally known techniques (e.g., liquidpeptide synthesis, solid phase peptide synthesis, convergent of smallpeptide fragments, etc.), and the synthesis method is not limited. Forexample, the OTP3 peptide of the present invention can be synthesizedusing the convergent method of small peptide fragments, in which sitesof a long-chain peptide to which a coupling can easily be made arevirtually cleaved and various parts are prepared based on this, and thencombined with one another to thereby finally synthesize a desiredpeptide. The above convergent method has limitations in that specificamino acids must exist within the peptide sequence, and thus, thepeptide may also be effectively synthesized by a combinatory peptidesynthesis method, where the solution phase synthesis method and thesolid phase peptide synthesis method are appropriately combined.

Experimental Example 1.2 Confirmation of Prepared Peptide 1—Analysis ofPurity

The purity of the peptide prepared in Experimental Example 1.1 wasmeasured by performing HPLC analysis (Shimadzu HPLC LabSolutions) usinga C-18 reversed-phase column (SHIMADZU C18 analytical column). As forthe analysis conditions, the sample was separated and developed into anaqueous solution of 0.05% trifluoroactate (TFA) and 0.05% TFA acetotrilesolution at 60° C., and then the purity was confirmed by measuring thepeak absorbance at a wavelength of 230 nm.

Experimental Example 1.3 Confirmation of Prepared Peptide 2—Analysis ofMolecular Weight

The molecular weight of the peptide prepared in Experimental Example 1.1was analyzed with a mass spectrometer (AXIMA Assurance, MALDI-TOF,Shimadzu).

Experimental Example 1.4 Confirmation of Prepared Peptide 2—Analysis ofQuantification

The peptide prepared in Experimental Example 1.1 was quantified bymeasuring the UV extinction coefficient (Ultrospec 3000 Pro UV/VISspectrophotometer, Pharmacia). Specifically, the quantificaiton wasperformed using the extinction coefficient at 280 nm.

Experimental Example 1.5 Preparation of Composition for In VivoAdministration

A composition for in vivo administration was prepared by mixing thepeptide prepared in Experimental Example 1.1, alhydrogel (Al(OH)₃,manufactured by InvivoGe, Inc.), and PHAD (manufactured by Avanti). Thespecific process is as follows.

(1) The prepared peptide powder was dissolved in 100% dimethyl sulfoxide(DMSO) to obtain a concentration of 100 mg/L.

(2) PBS was added to the peptide-DMSO solution of (1) and mixed toprepare the peptide at a concentration of 50 mg/mL.

(3) PHAD was dissolved in 100% DMSO to a concentration of 10 mg/mL, andthen diluted with distilled water to a concentration of 1 mg/mL.

(4) An alhydrogel adjuvant (Invivogen, USA) and the PHAD solution wereadded to the mixture of (2). The concentration of the mixed compositionwas 50 μg for the peptide, 10 μg for the PHAD, and 10% (v/v) for theAlhydrogel adjuvant, per 100 μL, which is the amount for one dose.

(5) After well mixing the mixture of (4), the resultant was reactedovernight while stirring with a rotator in a low temperature room (4°C.).

(6) For DSMO washing, the reactants of (5) were centrifuged at 1,400 rpmfor 15 minutes, and the supernatant except for about 1 mL above thepellet was removed. Thereafter, 10 mL of PBS was added thereto andmixed.

(7) The washing process of (6) was repeated 3 times.

(8) After the final washing process, PBS was added thereto so as toadjust the final concentration of the composition. In particular, thepeptide concentration was 50 μg/100 μL (in particular, 30 μg/100 μL inExample 4), the PHAD concentration was 10 μg/100 μL, and thelhydrogeladjuvant concentration was 10% (v/v).

(9) The amount of peptide adsorbed to the aluminum gel was measuredaccording to the method described in Experimental Example 2.3. As aresult, it was confirmed that the adsorption rate was 95% or more andthe peptide was used in the experiment.

Experimental Example 1.6 Preparation of Test Subject

In order to test the effect of the composition for in vivoadministration prepared in Experimental Example 1.5, C57BL/6, Balbcand/or ICR species were used as experimental mice (purchased fromCentral Lab Animal Inc.). Although there were some differences inspecies, the mice purchased were 7-week-old on average and they wereacclimatized for one week, and were used for experiments when they were8 weeks of age. Experimental mice were reared under the conditions of aconstant temperature and humidity environment within the temperaturerange of 23±1° C., relative humidity of 50±5%, and an environmentcontrolled as 12-hour light room/12 hour dark room. Drinking water andfood were provided ad libitum. In the case of a normal diet (purchasedfrom Central Lab Animal Inc.), it consisted of 20% protein, 70%carbohydrate, and 10% fat based on total calories, whereas in the caseof an obesity-inducing diet (purchased from Research diets), itconsisted of a high-fat diet including 20% protein, 20% carbohydrate,and 60% fat based on total calories. Experimental mice were divided intoeach group to have different dietary conditions and compositionadministration conditions, and experiments were performed with thenumber of individuals that could be statistically processed for eachexperimental group. Detailed conditions are as disclosed in eachspecific experimental example.

Experimental Example 1.7 Administration of Composition Including Peptideto Test Subject

For a test subject prepared in Experimental Example 1.6, differentcompositions were administered for each experimental group. Alladministration compositions were administered by an intramuscularinjection method, and after disinfecting muscles of both thighs of eachmouse with an alcohol swab, 50 μL each with a total amount of 100 μL wasinjected.

Experimental Example 1.8 Confirmation of Effect of Composition for InVivo Administration 1—Weight Measurement

In order to confirm the weight loss effect on a test subject of thecomposition administered in the body according to Experimental Example1.7, body weight and organ weight of the mice were measured for eachexperimental group. From the time of arrival to the end of theexperiment for each experimental group of mice, the average wascalculated after measuring three times per week to obtain the averageweight value for each week. After completion of the experiment, eachmouse was anesthetized, organs were dissected, and weights weremeasured, and the average was obtained for each experimental group.

Experimental Example 1.9 Confirmation of Effect of Composition for InVivo Administration 2—Confirmation of Antibody Titer

In order to determine whether the composition for in vivo administrationadministered in Experimental Example 1.7 induced an antibody against theB-cell epitope, a method for confirming the antibody titer using thetarget antigen is as follows. In particular, the target antigen isRNVPPIFNDVYWIAF (SEQ ID NO: 6) or ApoB100. The time to confirm theantibody titer may vary as needed, and the time was specificallydescribed for each experimental example.

1. Process of Antigen Coating Reaction

1-1) For each injection of a composition for in vivo administration intoa test subject, collect about 200 μL of blood from the subject's tailvein one week after the injection.

1-2) After placing the collected blood at 4° C. for one hour, performcentrifugation of the blood sample at 14,000 rpm for 10 minutes toseparate the serum, which is the supernatant.

1-3) Dilute the target antigen to a concentration of 50 μg/100 μL incoating buffer (0.05 M, bicarbonate, pH 9.6) and add the antigen to a96-well plate in an amount of 50 μg per well, and allow them to reactovernight at 4° C. so that the peptide is coated on the wall of thewell.

1-4) Wash the target antigen-coated plate three times using 300 μL ofphosphate buffered saline (PBS)-T (0.05% with Tween-20) per well.

2. Process of Blocking Reaction

2-1) Add 300 μL of a 0.5% casein blocking solution per well in theplate, and allow to react overnight at 4° C.

2-2) Wash the plate three times with 300 μL of PBS-T per well.

3. Process of Primary Antibody Reaction

3-1) As the primary antibody reaction, dilute the separated serum to anappropriate concentration, add it in an amount of 100 μL per well, andallow to react at 37° C. for one hour. In particular, after the initialinjection, subject the serum to serial dilution by diluting the same to1/20 to 1/1,000 in the experiment, and 1/500 to 1/10,000 depending onthe subject during the experiment after the second to third injections.As a positive control, use a monoclonal antibody against the peptide ofSEQ ID NO: 41 after purification.

3-2) After the reaction of 3-1, wash the plate three times with 300 μLof PBS-T per well.

4. Process of Secondary Antibody Reaction

4-1) As a secondary antibody reaction, add 100 μL of horseradishperoxidase (HRP) conjugated to anti-mouse IgG antibody recognizing amouse antibody per well, and allow them to react at 37° C. for one hour.

4-2) After the reaction of 4-1), wash the plate three times with 300 μLof PBS-T per well.

5. Process of Confirming Color Development and Absorbance

5-1) Add 100 μL of a o-phenylenediamine dihydrochloride (OPD) solutionper well, and allow to react at 37° C. for 10 minutes, and measure theabsorbance at OD 450 nm (Synergy HT microplate reader, BioTek).

Obtain the antibody titer in serum by converting the extinctioncoefficient, which is measured based on 1 mg/mL concentration of themonoclonal antibody against the target antibody as a positive control.

Experimental Example 1.10 Confirmation of Effect of Composition for InVivo Administration 3—Confirmation of Blood Lipid Concentration

The experimental method for confirming the effect of the composition forin vivo administration administered in Experimental Example 1.7 on theblood lipid concentration of a test subject is as follows:

(1) One week after the administration of each composition, collect about200 μL of blood from a subject's tail vein.

(2) Measurement of blood triglyceride (TG) concentration: Triglyzyme-V(Shinyak Chemical Co., Ltd.) was used. i) After mixing 4 μL of the bloodsample and 300 μL of a color developing reagent, the mixture was reactedat 37° C. for 5 minutes. ii) For the red quinone produced, theabsorbance was measured at 505 nm, and the concentration was calculatedby comparing with that of the reference solution.

(3) Measurement of total cholesterol concentration in blood:Cholestezyme-V (Shinyak Chemical Co., Ltd.) was used. i) After mixing 4μL of the blood sample and 300 μL of a color developing reagent, themixture was reacted at 37° C. for 5 minutes. ii) For the red quinoneproduced, the absorbance was measured at 505 nm, and the concentrationwas calculated by comparing with that of the reference solution.

(4) Measurement of high density lipoprotein (HDL) concentration inblood: HDL-C555 (Shinyak Chemical Co., Ltd.) was used. i) After mixing10 μL of the blood sample and 10 μL of a precipitation reagent, themixture was allowed to react at room temperature for at least 10minutes. ii) The reactants were centrifuged at 300 rpm or greater, andthe supernatant was separated. iii) After mixing 4 μL of the supernatantand 300 μL of a color developing reagent, the mixture was reacted at 37°C. for 5 minutes. iv) For the above reaction, the absorbance wasmeasured at 555 nm and the concentration was calculated by comparingwith that of the reference solution.

(5) Measurement of low density lipoprotein (LDL) concentration in blood.i) The reaction was performed using the Direct LDL Cholesterol detectionkit (Randox). ii) After the reaction of Step 2, for the quinoneproduced, the absorbance was measured at 600 nm, and the concentrationwas calculated by comparing with that of the reference solution.

Experimental Example 1.11 Confirmation of Effect of Composition for InVivo Administration 4—Confirmation of Lipolysis Ability and Comparisonof Size of Adipocytes

The method for confirming the effect of the composition for in vivoadministration administered in Experimental Example 1.7 on the abilityof degrading adipocytes and the size of adipocytes of a test subject byHormon sensitive lipase (HSL) is as follows:

1. Separation of Adipocytes

1-1) Cut an epididymal fat pad with scissors, add 4 mL per 1 g of KRBbuffer including 2% FBS, 2 mM glucose, and 1 mg/mL collagenase, shakeand allow them to react at 37° C. for one hour while shaking.

1-2) After the reaction is complete, pass the resultant through a 300 μmnylon mesh to filter out adipose tissue residues and adipose tissue, andthen pass the filtrate again through a 40 μm nylon mesh to separateadipocytes and macrophages.

1-3) Wash the adipocytes filtered in 1-2) by adding DMEM with 10% FBSand 1% AA, and remove the liquid in the lower layer with a syringe toobtain adipocytes from which collagenase has been removed.

2. Comparison of Lipolysis Ability

2-1) Seed the adipocytes obtained in 1-3) in a 48-well plate at 1.0×10⁵cells/well, and add a total of 1 mL of DMEM (10% FBS, 1% AA) thereto forculturing under 37° C. 5% CO₂ for two hours.

2-2) For the wells to induce HSL activity, add norepinephrine to a finalconcentration of 10⁻⁵M.

2-3) After the reaction is complete, allow 100 μL of the supernatant ofeach well to react with 100 μL of a free glycerol reagent, and measurethe absorbance at 540 nm.

3. Observation of Size of Adipocytes

3-1) Seed the adipocytes obtained in 1-3) in a 48 well plate at 1.0×10⁵cells/mL per well, treat with 10 μM of DAPI, allow them to react for twohours, and observe under a microscope.

3-2) In order to confirm whether DAPI-stained cells are adipocytes,stain lipids and nuclei together and observe. In particular, treat thecells with 10 μM of DAPI and 1:1,000 of HCS LipidTOX and allow them toreact for 24 hours, and then observe under a microscope.

Experimental Example 2 Confirmation of Peptide Effect 1 ExperimentalExample 2.1 Preparation of Peptides and Experiments Thereof

After preparing peptides according to [Table 1] according toExperimental Example 1.1, the peptides prepared were confirmed accordingto Experimental Examples 1.2 to 1.4. A composition for in vivoadministration including the peptides according to [Table 1] wasprepared, according to Experimental Example 1.5.

TABLE 1 Peptides used in Experimental Example 2 and compo-sition for in vivo administration including the same SEQ ID LabelPeptide sequence NO Example. 1 RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZ 56Example. 2 ZaK(Cha)VAAWTLKAAaZRNVPPIFNDVYWIAF 57

The test subject shown in [Table 2] was prepared according toExperimental Example 1.6.

TABLE 2 Test subject used in Experimental Example 2 Label Species DietComposition n Group 1-1 C57BL/6J, male ~13 weeks: normal diet Example. 16 after 13 weeks: high-fat diet Group 1-2 C57BL/6J, male ~13 weeks:normal diet Example. 2 6 after 13 weeks: high-fat diet

According to Experimental Example 1.7, the composition for in vivoadministration was administered to the test subject. In particular, theadministration cycle was as follows: 7 weeks, 9 weeks, 12 weeks, 15weeks, and 18 weeks of age.

Experimental Example 2.2 Confirmation of Experimental Results

In order to confirm the experimental results of Experimental Example2.1, the weight of the test subject was measured for each experimentalgroup disclosed in [Table 2] according to Experimental Example 1.8.

The experimental results are shown in FIGS. 1 to 3 .

Experimental Example 3 Confirmation of Peptide Effect 2 ExperimentalExample 3.1 Peptide Preparation and Experiment Thereof

After preparing the peptides according to [Table 3] according toExperimental Example 1.1, the peptides prepared were confirmed accordingto Experimental Examples 1.2 to 1.4. A composition for in vivoadministration including the peptides according to [Table 3] accordingto Experimental Example 1.5 was prepsared.

TABLE 3 Peptides used in Experimental Example 3 and compo-sition for in vivo administration including the same SEQ ID LabelPeptide sequence NO Example. RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAa 108 3ZCRFRGLISLSQVYLSThe test subject shown in [Table 4] was prepared according toExperimental Example 1.6.

TABLE 4 Test subject used in Experimental Example 3 Label Species DietComposition n Lean C57BL/6J, male ~10 weeks: normal diet PBS-Alum 12after 10 weeks: high-fat diet Obesity C57BL/6J, male ~10 weeks: normaldiet PBS-Alum 12 after 10 weeks: high-fat diet Mock C57BL/6J, male ~10weeks: normal diet Alum-PHAD/ 12 after 10 weeks: high-fat PBS diet Group2-1 C57BL/6J, male ~10 weeks: normal diet Example. 3 12 (3H-OTP) after10 weeks: high-fat diet

In particular, Lean denotes a control group with normal weight, Obesedenotes an obesity group induced by high-fat diet, and Mock denotes agroup administered with placebo (the same hereinafter).

According to Experimental Example 1.7, the composition for in vivoadministration was administered to the test subject. In particular, theadministration cycle was as follows: 8 weeks, 10 weeks, 12 weeks, and 14weeks of age.

Experimental Example 3.2 Confirmation of Experimental Results

In order to confirm the experimental results of Experimental Example3.1, the following experiments were performed in a test subject for eachexperimental group disclosed in [Table 4].

(1) According to Experimental Example 1.8, the weight of a test subjectfor each experimental group was measured.

(2) According to Experimental Example 1.9, the antibody titers observedin a test subject for each experimental group were confirmed.

(3) According to Experimental Example 1.10, the blood lipidconcentration of a test subject for each experimental group wasmeasured.

(4) According to Experimental Example 1.11, the lipolysis ability of atest subject for each experimental group was confirmed, and the size ofadipocytes was observed.

The experimental results are shown in FIGS. 4 to 9 .

Experimental Example 4 Confirmation of Peptide Effect 3 ExperimentalExample 4.1 Peptide Preparation and Experiment Thereof

After preparing the peptides according to [Table 5] according toExperimental Example 1.1, the peptides prepared were confirmed accordingto Experimental Examples 1.2 to 1.4. A composition for in vivoadministration including the peptides according to [Table 5] accordingto Experimental Example 1.5 was prepared.

TABLE 5 Peptides used in Experimental Example 4 and compo-sition for in vivo administration including the same SEQ ID LabelPeptide sequence NO Example. RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAa 108 3ZCRFRGLISLSQVYLS Example. RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAa 108 4ZCRFRGLISLSQVYLS

In particular, in Example.4, the final peptide concentration was 30μg/100 μL.

The test subject shown in [Table 6] was prepared according toExperimental Example 1.6.

TABLE 6 Test subject used in Experimental Example 4 Label Species DietComposition Lean C57BL/6J, male ~11 weeks: normal diet PBS-Alum 8 after11 weeks: high-fat diet Obesity C57BL/6J, male ~11 weeks: normal dietPBS-Alum 9 after 11 weeks: high-fat diet Mock C57BL/6J, male ~11 weeks:normal diet Alum-PHAD/ 9 after 11 weeks: high-fat PBS diet Group 3-1C57BL/6J, male ~11 weeks: normal diet Example. 3 8 (3H-OTP after 11weeks: high-fat 50 ug) diet Group 3-2 C57BL/6J, male ~11 weeks: normaldiet Example. 4 8 (3H-OTP after 11 weeks: high-fat 30 ug) diet

According to Experimental Example 1.7, the composition for in vivoadministration was administered to the test subject. In particular, theadministration cycle was as follows: 8 weeks, 11 weeks, 14 weeks, 17weeks, and 20 weeks of age.

Experimental Example 4.2 Confirmation of Experimental Results

In order to confirm the experimental results of Experimental Example4.1, the following experiments were performed in a test subject for eachexperimental group disclosed in [Table 6].

(1) According to Experimental Example 1.8, the weight of a test subjectfor each experimental group was measured.

(2) According to Experimental Example 1.9, the antibody titers observedin a test subject for each experimental group were confirmed.

The experimental results are shown in FIGS. 10 and 11 .

Experimental Example 5 Confirmation of Peptide Effect 4 ExperimentalExample 5.1 Peptide Preparation and Experiment Thereof

After preparing the peptides according to [Table 7] according toExperimental Example 1.1, the peptides prepared were confirmed accordingto Experimental Examples 1.2 to 1.4. A composition for in vivoadministration including the peptides according to [Table 7] accordingto Experimental Example 1.5 was prepared.

TABLE 7 Peptides used in Experimental Example 5 and compo-sition for in vivo administration including the same SEQ ID LabelPeptide sequence NO Example. RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAa 108 3ZCRFRGLISLSQVYLS

The test subject shown in [Table 8] was prepared according toExperimental Example 1.6.

TABLE 8 Test subject used in Experimental Example 5 Label Species DietComposition n Lean C57BL/6J, male normal diet PBS-Alum 10 ObesityC57BL/6J, male ~5 weeks: normal diet PBS-Alum 10 after 5 weeks: high-fatdiet Group 4-1 C57BL/6J, male ~5 weeks: normal diet Example. 3 10(3H-OTP) after 5 weeks: high-fat diet

According to Experimental Example 1.7, the composition for in vivoadministration was administered to the test subject. In particular, theadministration cycle was as follows: 11 weeks, 13 weeks, 15 weeks, and17 weeks of age.

Experimental Example 5.2 Confirmation of Experimental Results

In order to confirm the experimental results of Experimental Example5.1, the weight of the test subject was measured for each experimentalgroup disclosed in [Table 8] according to Experimental Example 1.8.

The experimental results are shown in FIG. 12 .

Experimental Example 6 Confirmation of Peptide Effect 5 ExperimentalExample 6.1 Peptide Preparation and Experiment Thereof

After preparing the peptides according to [Table 9] according toExperimental Example 1.1, the peptides prepared were confirmed accordingto Experimental Examples 1.2 to 1.4. A composition for in vivoadministration including the peptides according to [Table 9] accordingto Experimental Example 1.5 was prepared.

TABLE 9 Peptides used in Experimental Example 6 and compo-sition for in vivo administration including the same SEQ ID LabelPeptide sequence NO Example. RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAa 108 3ZCRFRGLISLSQVYLS

The test subject shown in [Table 10] was prepared according toExperimental Example 1.6.

TABLE 10 Test subject used in Experimental Example 6 Label Species DietComposition n Lean C57BL/6J- normal diet PBS-Alum Female: 3 Rag2em1hwl/Korl, female/male Wild(+/+) Group 5-1 C57BL/6J- ~10 weeks: normalPBS-Alum Female: 4 (Wild(+/+)) Rag2em1hwl/ diet Male: 3 Korl, after 10weeks: female/male high-fat diet Wild(+/+) Group 5-2 C57BL/6J- ~10weeks: normal Example. 3 Female: 3 (Hetero(+/−)) Rag2em1hwl/ diet Male:3 Korl, after 10 weeks: female/male high-fat diet Hetero(+/−) Group 5-3C57BL/6J- ~10 weeks: normal Example. 3 Female: 3 (Homo(−/−)) Rag2em1hwl/diet Male: 1 Korl, after 10 weeks: female/male high-fat diet Homo(−/−) *The C57BL/6J-Rag2em1hwl/Korl mouse is a mouse in which the Rag2 gene,which is a gene involved in the antibody producing ability in a C57BL/6Jmouse, is knocked out. ** Wild (+/+) refers to a wild-type mouse inwhich no gene is mutated, Hetero (+/−) refers to a heterozygote mouse,and Homo (−/−) refers to a homozygote mouse. *** Homo (−/−) mouse doesnot have the ability to produce antibodies in the body.

According to Experimental Example 1.7, the composition for in vivoadministration was administered to the test subject. In particular, theadministration cycle was as follows: 8 weeks, 10 weeks, 12 weeks, and 14weeks of age.

Experimental Example 6.2 Confirmation of Experimental Results

In order to confirm the experimental results of Experimental Example6.1, the weight of the test subject was measured for each experimentalgroup disclosed in [Table 10] according to Experimental Example 1.8.

The experimental results are shown in FIG. 13 .

Experimental Example 7 Confirmation of Peptide Effect 6 ExperimentalExample 7.1 Peptide Preparation and Experiment Thereof

After preparing the peptides according to [Table 11] according toExperimental Example 1.1, the peptides prepared were confirmed accordingto Experimental Examples 1.2 to 1.4. A composition for in vivoadministration including the peptides according to [Table 11] accordingto Experimental Example 1.5 was prepared.

TABLE 11 Peptides used in Experimental Example 7 and compo-sition for in vivo administration including the same SEQ ID LabelPeptide sequence NO Example. RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAa 108 5ZCRFRGLISLSQVYLS Example. RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAa 68 6ZGSHHHHHHGSDDDDK Example. GSHHHHHHGSDDDDKZaK(Cha)VAAWTLKAAa 69 7ZRNVPPIFNDVYWIAF Example. KTTKQSFDLSVKAQYKKNKHZaK(Cha)VAAWT 114 8LKAAaZCRFRGLISLSQVYLS Example. RNVPPIFNDVYWIAFCRFRGLISLSQVYLSZaK 84 9(Cha)VAAWTLKAAaZ Example. RNVPPIFNDVYWIAFZPKYVKQNTLKLATZCRF 121 10RGLISLSQVYLS Example. RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAa 56 11 Z Example.RNVPPIFNDVYWIAFK(Cha)VAAWTLKAA 62 12 Example.RNVPPIFNDVYWIAFK(Cha)VAAWTLKAAHHH 67 13 HHH

The test subject shown in [Table 12] was prepared according toExperimental Example 1.6.

TABLE 12 Test subject used in Experimental Example 7 Label Species DietComposition n Lean C57BL/6J, male ~10 weeks: normal diet PBS-Alum 8after 10 weeks: high-fat diet Obese C57BL/6J, male ~10 weeks: normaldiet PBS-Alum 12 after 10 weeks: high-fat diet Group 6-1 C57BL/6J, male~10 weeks: normal diet Example. 5 12 (P1) after 10 weeks: high-fat dietGroup 6-2 C57BL/6J, male ~10 weeks: normal diet Example. 6 8 (P2) after10 weeks: high-fat diet Group 6-3 C57BL/6J, male ~10 weeks: normal dietExample. 7 8 (P3) after 10 weeks: high-fat diet Group 6-4 C57BL/6J, male~10 weeks: normal diet Example. 8 8 (P4) after 10 weeks: high-fat dietGroup 6-5 C57BL/6J, male ~10 weeks: normal diet Example. 9 4 (P5) after10 weeks: high-fat diet Group 6-6 C57BL/6J, male ~10 weeks: normal dietExample. 10 8 (P6) after 10 weeks: high-fat diet Group 6-7 C57BL/6J,male ~10 weeks: normal diet Example. 11 8 (P7) after 10 weeks: high-fatdiet Group 6-8 C57BL/6J, male ~10 weeks: normal diet Example. 12 8 (P8)after 10 weeks: high-fat diet Group 6-9 C57BL/6J, male ~10 weeks: normaldiet Example. 13 8 (P9) after 10 weeks: high-fat diet

According to Experimental Example 1.7, the composition for in vivoadministration was administered to the test subject. In particular, theadministration cycle was as follows: 8 weeks, 10 weeks, 12 weeks, and 14weeks of age.

Experimental Example 7.2 Confirmation of Experimental Results

In order to confirm the experimental results of Experimental Example7.1, the following experiments were performed in a test subject for eachexperimental group disclosed in [Table 12].

(1) According to Experimental Example 1.8, the weight of a test subjectfor each experimental group was measured.

(2) According to Experimental Example 1.9, the antibody titers observedin a test subject for each experimental group at 11 weeks, 16 weeks, and19 weeks were confirmed. In particular, the target antigen wasdetermined using RNVPPIFNDVYWIAF (SEQ ID NO: 6).

The experimental results are shown in FIGS. 14 to 25 .

As a result of the experiment, all of the experimental groups 6-1 (P1)to experimental groups 6-9 (P9) showed a distinct weight loss effectcompared to the control group (Obese), which were fed with a high-fatdiet.

The results of the antibody titer test according to the experimentalgroup can be interpreted as follows:

Since no antigen was administered to the control groups (i.e., Lean andObese), antibody titers were not shown.

In consideration of the experimental design, in the experimental groups,the antibody titers to SEQ ID NO: 6 included in the peptides used weremainly confirmed. Although antibody titers to sequences other than SEQID NO: 6 included in the peptides used were not separately confirmed,the weight loss effect due to a humoral immunity by these was confirmedtogether.

Specific results will be described below.

In the case of Experimental Group 6-1 (P1), there were individualsshowing a large antibody titer depending on the individual, and therewere also individuals which showed no antibody titer. These results canbe interpreted as follows: for some individuals of the experimentalgroup 6-1 (P1), a humoral immunity to RNVPPIFNDVYWIAF (SEQ ID NO: 6)included in Example 6 was induced, thus showing both the weight losseffect and antibody titer observed; and although for some individuals, ahumoral immunity to CRFRGLISLSQVYLS (SEQ ID NO: 7) included in Example 6was induced, resulting in weight loss, the antigen used in theEnzyme-Linked Immunosorbent Assay (ELISA) consisted only ofRNVPPIFNDVYWIAF (SEQ ID NO: 6) and there was no other antigen (i.e., theantigen of SEQ ID NO: 7), therefore, the antibody titer according to theabove experiment was not observed even in the presence of the antibody.

In the case of experimental group 6-2 (P2), the antibody titer toRNVPPIFNDVYWIAF (SEQ ID NO: 6) included in Example 7 was observed, andthe weight loss effect appeared, it can be interpreted that the B-cellepitope included in the peptide of Example 7 well induced a humoralimmunity thereto.

In the case of experimental group 6-3 (P3), the antibody titer toRNVPPIFNDVYWIAF (SEQ ID NO: 6) included in Example 8 was observed, andthe weight loss effect appeared, it can be interpreted that that theB-cell epitope included in the peptide of Example 8 well induced ahumoral immunity thereto.

In the case of experimental group 6-4 (P4), it can be interpreted that ahumoral immunity to KTTKQSFDLSVKAQYKKNKH (SEQ ID NO: 8) and/orCRFRGLISLSQVYLS (SEQ ID NO: 7) included in Example 9 was induced, thusantibody titers against RNVPPIFNDVYWIAF (SEQ ID NO: 6) were not shown,while there was a weight loss effect.

These results can be interpreted that for some individuals of theexperimental group 6-5 (P5), a humoral immunity was induced againstRNVPPIFNDVYWIAF (SEQ ID NO: 6) included in Example 10, such that both aweight loss effect and an antibody titer are observed, for the otherindividuals of the experimental group 6-5 (P5), a humoral immunity wasinduced against CRFRGLISLSQVYLS (SEQ ID NO: 7) included in Example 10,such that a weight loss effect is observed, but the antibody titeraccording to the above experiment was not observed.

In the case of experimental group 6-7 (P7), although there were somedifferences between individuals, antibodies to RNVPPIFNDVYWIAF (SEQ IDNO: 6) was observed and the weight loss effect appeared, therefore, itcan be interpreted that the B-cell epitope included in the peptide ofExample 11 well induced a humoral immunity.

In the case of experimental group 6-8 (P8), since the antibody titeragainst RNVPPIFNDVYWIAF (SEQ ID NO: 6) included in Example 12 was shown,and the weight loss effect was shown, it can be interpreted that theB-cell epitope included in the peptide of Example 12 well induced ahumoral immunity.

In the case of experimental group 6-9 (P9), there were some differencesbetween individuals. However, since the antibody against RNVPPIFNDVYWIAF(SEQ ID NO: 6) was observed and the weight loss effect appeared, it canbe interpreted that the B-cell epitope included in the peptide ofExample 13 well induced a humoral immunity.

Experimental Example 8 Confirmation of Peptide Effect 7 ExperimentalExample 8.1 Peptide Preparation and Experiment Thereof

After preparing the peptides according to [Table 13] according toExperimental Example 1.1, the peptides prepared are confirmed accordingto Experimental Examples 1.2 to 1.4. A composition for in vivoadministration including the peptides according to [Table 13] accordingto Experimental Example 1.5 are prepared.

TABLE 13 Peptides used in Experimental Example 8 and compo-sition for in vivo administration including the same SEQ ID LabelPeptide sequence NO Example. RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAa 108 5SZCRFRGLISLQVYLS Example. RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAC 160 14RFRGLISLSQVYLS Example. RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAACR 161 15

The test subject shown in [Table 14] is prepared according toExperimental Example 1.6.

TABLE 14 Test subject used in Experimental Example 8 Label Species DietComposition n Lean C57BL/6J, male ~10 weeks: normal diet PBS-Alum 8after 10 weeks: high-fat diet Obese C57BL/6J, male ~10 weeks: normaldiet PBS-Alum 8 after 10 weeks: high-fat diet Group 7-1 C57BL/6J, male~10 weeks: normal diet Example. 5 8 after 10 weeks: high-fat diet Group7-2 C57BL/6J, male ~10 weeks: normal diet Example. 14 8 after 10 weeks:high-fat diet Group 7-3 C57BL/6J, male ~10 weeks: normal diet Example.15 8 after 10 weeks: high-fat diet

According to Experimental Example 1.7, the composition for in vivoadministration is administered to the test subject. In particular, theadministration cycle, administration timing, and frequency ofadministration may be appropriately modified according to theexperimental design. For example, the composition may be administeredfour times at intervals of two weeks for a test subject of 8 weeks ofage, but is not limited thereto.

The above experimental method may be appropriately modified asnecessary.

Experimental Example 8.2 Confirmation of Experimental Results

In order to confirm the experimental results of Experimental Example8.1, the following experiments are performed in a test subject for eachexperimental group disclosed in [Table 14].

(1) According to Experimental Example 1.8, the weight of a test subjectfor each experimental group is measured.

(2) According to Experimental Example 1.9, the antibody titers observingin a test subject for each experimental group are confirmed.

(3) According to Experimental Example 1.10, the blood lipidconcentration of a test subject for each experimental group is measured.

(4) According to Experimental Example 1.11, the lipolysis ability of atest subject in each experimental group is confirmed and the size ofadipocytes is observed.

Experimental Example 9 Confirmation of Peptide Effect 8 ExperimentalExample 9.1 Peptide Preparation and Experiment Thereof

According to Experimental Example 1.1, a peptide represented by one ormore sequences selected from the group consisting of the following isprepared: SEQ ID NOS: 56 to 158, SEQ ID NOS: 160 to 161, and SEQ ID NOS:198 to 220.

The peptides prepared are confirmed according to Experimental Examples1.2 to 1.4. According to Experimental Example 1.5, a composition for invivo administration including the peptides prepared is prepared.

In particular, the peptide may be prepared by selecting only somesequences from the sequence groups above, and repeated experiments maybe performed with multiple combinations as needed.

The test subject for the peptides prepared is prepared according toExperimental Example 1.6. In particular, examples of the control groupsand the experimental groups to be used are as shown in [Table 15], andeach experimental group is determined with reference to the conditionsof Experimental Example 1.6 and [Table 15].

TABLE 15 Test subject used in Experimental Example 9 Label Species DietComposition n Lean C57BL/6J, male ~10 weeks: normal diet PBS-Alum 8after 10 weeks: high-fat diet Obese C57BL/6J, male ~10 weeks: normaldiet PBS-Alum 8 after 10 weeks: high-fat diet Experimental C57BL/6J,male ~10 weeks: normal diet One of the 8 group after 10 weeks: high-fatprepared diet Example in Experimental example 9.1

In particular, the experimental groups are prepared as many as thenumber of the prepared peptides prepared.

According to Experimental Example 1.7, the composition for in vivoadministration is administered to the test subject. In particular, theadministration cycle, administration timing, and frequency ofadministration may be appropriately modified according to theexperimental design. For example, the composition may be administeredfour times at intervals of two weeks for a test subject of 8 weeks ofage, but is not limited thereto.

The above experimental method may be appropriately modified asnecessary.

Experimental Example 9.2 Confirmation of Experimental Results

In order to confirm the experimental results of Experimental Example9.1, the following experiments are performed in a test subject for eachexperimental group.

(1) According to Experimental Example 1.8, the weight of a test subjectfor each experimental group is measured.

(2) According to Experimental Example 1.9, the antibody titers observingin a test subject for each experimental group are confirmed.

(3) According to Experimental Example 1.10, the blood lipidconcentration of a test subject for each experimental group is measured.

(4) According to Experimental Example 1.11, the lipolysis ability of atest subject in each experimental group was confirmed and the size ofadipocytes is observed.

Experimental Example 10 Confirmation of Peptide Effect 9 ExperimentalExample 10.1 Peptide Preparation and Experiment Thereof

After preparing the peptides according to Examples 81 to 90 in theparagraph of “Possible Examples of the Invention” according toExperimental Example 1.1, the peptides prepared are confirmed accordingto Experimental Examples 1.2 to 1.4. According to Experimental Example1.5, a composition for in vivo administration including the peptidesprepared is prepared.

In particular, only parts of the peptides according to Examples 81 to 90of the paragraph of “Possible Examples of the Invention” may be selectedand prepared, and experiments on multiple combinations may be repeatedas needed.

The test subject for the peptide prepared is prepared according toExperimental Example 1.6. In particular, the control groups andexperimental groups to be used are determined with reference to theconditions of Experimental Example 1.6 and [Table 15].

According to Experimental Example 1.7, the composition for in vivoadministration is administered to the test subject. In particular, theadministration cycle, administration timing, and frequency ofadministration may be appropriately modified according to theexperimental design. For example, the composition may be administeredfour times at intervals of two weeks for a test subject of 8 weeks ofage, but is not limited thereto.

The above experimental method may be appropriately modified asnecessary.

Experimental Example 10.2 Confirmation of Experimental Results

In order to confirm the experimental results of Experimental Example10.1, the following experiments are performed in a test subject for eachexperimental group.

(1) According to Experimental Example 1.8, the weight of a test subjectfor each experimental group is measured.

(2) According to Experimental Example 1.9, the antibody titers observingin a test subject for each experimental group are confirmed.

(3) According to Experimental Example 1.10, the blood lipidconcentration of a test subject for each experimental group is measured.

(4) According to Experimental Example 1.11, the lipolysis ability of atest subject in each experimental group is confirmed and the size ofadipocytes is observed.

INDUSTRIAL APPLICABILITY

A peptide unit, a peptide, and/or a nucleic acid encoding the sameprovided herein can be used to prepare an immunotherapeutic,particularly an agent for treating obesity, and the immunotherapeuticmay show a therapeutic effect by generating an intended humoral immunitywhen administered into the body of a subject.

1-25. (canceled)
 26. A method for treating obesity comprisingadministering a peptide to a subject, wherein the peptide is representedby [Formula 1]:N-B-A₁-T-A₂-C;   [Formula 1] wherein, B is a sequence of B-cell epitopefor Apolipoprotein B-100, wherein the sequence of B-cell epitope forApolipoprotein B-100 is at least 90% identical to a sequence selectedfrom RNVPPIFNDVYWIAF (SEQ ID NO: 6), CRFRGLISLSQVYLS (SEQ ID NO: 7), andKTTKQSFDLSVKAQYKKNKH (SEQ ID NO: 8); wherein A₁ is a sequence of firstauxiliary part selected from Za and Z, or is absent; wherein T is asequence of Helper-T-cell epitope (Th epitope), wherein the sequence ofTh epitope is at least 90% identical to a sequence selected fromK(Cha)VAAWTLKAA (SEQ ID NO: 1), PKYVKQNTLKLAT (SEQ ID NO: 2),ILMQYIKANSKFIGI (SEQ ID NO: 3), QSIALSSLMVAQAIP (SEQ ID NO: 4),ILMQYIKANSKFIGIPMGLPQSIALSSLMVAQ (SEQ ID NO: 5), PLGFFPDHQL (SEQ ID NO:162), WPEANQVGAGAFGPGF (SEQ ID NO: 163), MQWNSTALHQALQDP (SEQ ID NO:164), MQWNSTTFHQTLQDPRVRGLYFPAGG (SEQ ID NO: 165), FFLLTRILTI (SEQ IDNO: 166), FFLLTRILTIPQSLD (SEQ ID NO: 167), TSLNFLGGTTVCLGQ (SEQ ID NO:168), QSPTSNHSPTSCPPIC (SEQ ID NO: 169), IIFLFILLLCLIFLLVLLD (SEQ ID NO:170), CTTPAQGNSMFPSC (SEQ ID NO: 171), CTKPTDGN (SEQ ID NO: 172),WASVRFSW (SEQ ID NO: 173), LLPIFFCLW (SEQ ID NO: 174),MDIDPYKEFGATVELLSFLP (SEQ ID NO: 175), FLPSDFFPSV (SEQ ID NO: 176),RDLLDTASALYREALESPEH (SEQ ID NO: 177), PHHTALRQAILCWGELMTLA (SEQ ID NO:178), GRETVIEYLVSFGVW (SEQ ID NO: 179), EYLVSFGVWIRTPPA (SEQ ID NO:180), VSFGVWIRTPPAYRPPNAPI (SEQ ID NO: 181), TVVRRRGRSP (SEQ ID NO:182), VGPLTVNEKRRLKLI (SEQ ID NO: 183), RHYLHTLWKAGILYK (SEQ ID NO:184), ESRLVVDFSQFSRGN (SEQ ID NO: 185), LQSLTNLLSSNLSWL (SEQ ID NO:186), SSNLSWLSLDVSAAF (SEQ ID NO: 187), LHLYSHPIILGFRKI (SEQ ID NO:188), KQCFRKLPVNRPIDW (SEQ ID NO: 189), LCQVFADATPTGWGL (SEQ ID NO:190), AANWILRGTSFVYVP (SEQ ID NO: 191), and EIRLKVFVLGGCRHK (SEQ ID NO:192), wherein A denotes D-form alanine, (Cha) denotesL-cyclohexylalanine, and the Z denotes 6-aminohexanoic acid; wherein A₂is a sequence of second auxiliary part selected from Z, CR, HHHHHH (SEQID NO: 53), and ZGSHHHHHHGSDDDDK (SEQ ID NO: 194), or is absent; whereinthe peptide length is 23mer to 71mer; and wherein the peptide isrecognized by CD4+ T-cell to induce a humoral immune response.
 27. Themethod of claim 26, wherein A1 of the peptide is Za, A2 of the peptideis aZGSHHHHHHGSDDDDK (SEQ ID NO: 194), and the B of the peptide isKTTKQSFDLSVKAQYKKNKH (SEQ ID NO: 8).
 28. The method of claim 27, whereinthe peptide is RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZGSHHHHHHGSDDDDK (SEQ IDNO: 68).
 29. The method of claim 26, wherein A₁ of the peptide is Za, A₂of the peptide is aZ, and B of the peptide is RNVPPIFNDVYWIAF (SEQ IDNO: 6).
 30. The method of claim 29, wherein the peptide isRNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAAaZ (SEQ ID NO: 56).
 31. The method ofclaim 26, wherein A₁ of the peptide is absent, A₂ of the peptide isHHHHHH (SEQ ID NO; 53), and B of the peptide is RNVPPIFNDVYWIAF (SEQ IDNO: 6).
 32. The method of claim 31, wherein the peptide isRNVPPIFNDVYWIAFK(Cha)VAAWTLKAAHHHHHH (SEQ ID NO: 67)
 33. The method ofclaim 26, wherein A₁ of the peptide is CR, and B of the peptide isRNVPPIFNDVYWIAF (SEQ ID NO: 6).
 34. The method of claim 33, wherein thepeptide is RNVPPIFNDVYWIAFZaK(Cha)VAAWTLKAACR (SEQ ID NO: 161).